_:b42012392 _:b42012407 . _:b42012392 _:b42012400 . . _:b42012392 _:b42012401 . _:b42012392 _:b42012402 . . _:b42012392 _:b42012403 . . _:b582918956 . _:b42012361 "SV40 large T-immortalized Lats1\u2013/\u2013Lats2F/F MEFs were described previously (Murakami et al. >>2011<<). For Lats2 deletion, MEFs were infected with an adenovirus encoding Cre recombinase (no." . . _:b582918473 . . . _:b582918999 . . . . . _:b582918488 . . . _:b582918762 . . . . _:b582918930 . _:b582918456 . _:b582918613 . . . _:b582918776 . . . _:b582918595 . . . _:b582918516 . _:b582918744 . _:b582918948 . . _:b42012388 . _:b582919184 . _:b582918773 . . _:b582919078 . . _:b582918917 . _:b582918987 . _:b582918425 . . . _:b582918745 . _:b582918829 . . . . _:b582919076 . _:b582918521 . _:b582918860 . . . _:b42012371 . . _:b582918661 "4"^^ . _:b582919138 . _:b582918660 "4"^^ . _:b582918663 "4"^^ . _:b582918907 . . _:b582918662 "4"^^ . . . _:b582918657 "4"^^ . . _:b582918656 "4"^^ . . _:b42012372 "Consistent with the observed LATS1 and LATS2 activation, phosphorylation of angiomotin (AMOT) at Ser175, a known direct substrate site for LATS1/2 kinases (Adler et al. 2013; Chan et al. 2013; Dai et al. >>2013<<; Hirate et al. 2013), was increased in TAZ(4SA)-overexpressing MCF10A cells (Fig." . _:b582918539 . . _:b582918659 "4"^^ . _:b582918658 "4"^^ . _:b582918669 "4"^^ . . . . _:b582918668 "4"^^ . _:b582919193 . _:b582918620 . _:b582918671 "4"^^ . _:b582919144 . . _:b42012378 "As we observed an increased phosphorylation of LATS1/2 at the HM, a site known to be phosphorylated by MST1/2 (Chan et al. >>2005<<), we investigated whether MST1/2 are responsible for this activation." . _:b582918670 "4"^^ . . _:b582918665 "4"^^ . . . . _:b582918664 "4"^^ . _:b582918969 . _:b582918667 "4"^^ . . _:b582918666 "4"^^ . . _:b582918677 "4"^^ . _:b582918676 "4"^^ . _:b582918679 "4"^^ . _:b582918540 . _:b582918678 "4"^^ . . . _:b582919111 . _:b582918673 "4"^^ . _:b582918672 "4"^^ . . . _:b582918675 "4"^^ . . _:b582918674 "4"^^ . _:b42012392 "discussion" . _:b582918474 . _:b582918685 "4"^^ . _:b582919210 . _:b582918684 "4"^^ . _:b582919196 . _:b582918687 "4"^^ . . _:b582918862 . _:b582918686 "4"^^ . . _:b582918681 "4"^^ . _:b582918680 "4"^^ . . _:b582918510 . _:b582918683 "4"^^ . . _:b582918682 "4"^^ . . _:b582918693 "3"^^ . _:b582918700 . _:b582918692 "3"^^ . _:b582918701 . _:b582918702 . _:b582918703 . _:b582918696 . _:b582918727 . _:b582918695 "3"^^ . _:b582919200 . _:b582918697 . . _:b582918698 . _:b582918699 . _:b582918692 . _:b582919028 . _:b582918694 "3"^^ . _:b582918693 . _:b582918694 . _:b582918695 . . _:b582918688 . _:b582918689 "3"^^ . _:b582918689 . . _:b582918883 . _:b582918690 . _:b582918691 . . _:b582918716 . _:b582918688 "3"^^ . _:b582919005 . _:b582918717 . _:b582918718 . . _:b582918719 . _:b582918712 . _:b582918691 "3"^^ . _:b582918713 . _:b582918714 . _:b582918715 . . . _:b582918708 . _:b582918690 "3"^^ . _:b582918709 . _:b582918710 . _:b582918711 . _:b582918704 . _:b582918701 "3"^^ . _:b582918705 . _:b582918659 . _:b582918706 . _:b582918707 . . _:b582918668 . _:b582918700 "3"^^ . _:b582918669 . . _:b582918670 . _:b582918671 . _:b582918946 . _:b582918664 . _:b582918703 "3"^^ . _:b582919145 . _:b582918665 . . . _:b582918666 . _:b582918667 . _:b582918660 . _:b582918702 "3"^^ . _:b582918821 . _:b582918661 . _:b582918662 . _:b582918663 . _:b582918656 . _:b582918697 "3"^^ . _:b582918657 . _:b582918658 . _:b582918659 . _:b582918684 . _:b582918423 . _:b582918685 . _:b582918696 "3"^^ . _:b582918686 . _:b582918625 . _:b582918687 . _:b582918680 . _:b582918699 "3"^^ . _:b582918681 . _:b582918682 . _:b582918683 . _:b582918676 . _:b42012387 "While YAP and TAZ are known to promote cell migration (Chan et al. 2008; Haskins et al. >>2014<<), recent studies revealed that the phosphorylation of AMOT by LATS1/2 inhibits AMOT binding to F-actin (Chan et al. 2013; Dai et al. 2013; Hirate et al. 2013), reduces actin stress fiber formation, and thereby inhibits migration (Dai et" . _:b582918698 "3"^^ . _:b582918677 . _:b582918640 . _:b582918678 . _:b582918679 . . _:b582918672 . _:b582918709 "3"^^ . _:b582918673 . . . _:b582918674 . _:b582918675 . _:b582918764 . _:b582918708 "3"^^ . _:b582918765 . _:b582918766 . _:b582918767 . _:b582918760 . _:b582918711 "3"^^ . _:b582918761 . _:b582918762 . _:b582918763 . _:b582918756 . _:b582918710 "3"^^ . _:b582918757 . _:b582918758 . _:b582918759 . _:b582918752 . _:b582918705 "3"^^ . _:b582919160 . _:b582918753 . . _:b582918754 . _:b582918755 . _:b582918780 . . _:b582918781 . _:b582918704 "3"^^ . _:b582918782 . . _:b582918783 . _:b582918776 . _:b582919097 . _:b582918707 "3"^^ . _:b582918777 . _:b582918778 . _:b582918779 . _:b582918772 . _:b582918706 "3"^^ . _:b582918773 . . _:b582918774 . _:b582918775 . _:b582918768 . _:b42012393 "The negative feedback mechanism employed by the mammalian Hippo pathway appears to differ significantly from that in D. melanogaster, where Yorkie induces expression of expanded, merlin, four-jointed, or kibra (Cho et al. >>2006<<; Hamaratoglu et al. 2006; Genevet et al. 2010), but not warts (the fly homolog of LATS1/2), in dividing epithelial cells of the wing or eye disc (Jukam et al. 2013)." . _:b582918717 "3"^^ . _:b582918769 . _:b582918770 . _:b582918771 . _:b582918732 . _:b582918716 "3"^^ . _:b582918733 . _:b582918734 . . _:b582918735 . _:b582918728 . _:b582918430 . _:b582918729 . _:b582918719 "3"^^ . _:b582918730 . _:b582918865 . _:b582918731 . _:b582918724 . _:b582919018 . _:b582918718 "3"^^ . _:b582918993 . _:b582918725 . _:b582918726 . . _:b582918727 . _:b582918720 . _:b582918629 . _:b582918721 . . _:b582918713 "3"^^ . . _:b582918722 . _:b582918723 . _:b582918748 . _:b582918712 "3"^^ . _:b582918749 . _:b582918750 . _:b582918751 . _:b582918744 . _:b582918715 "3"^^ . _:b582918745 . _:b582918746 . _:b582918747 . . _:b582918740 . . _:b582918741 . _:b582918714 "3"^^ . _:b582918742 . _:b582918743 . _:b582918736 . _:b582918771 . _:b582918725 "3"^^ . _:b42012396 . _:b582918737 . _:b582918453 . _:b582918738 . _:b582918739 . _:b582918828 . _:b582918724 "3"^^ . _:b42012397 . _:b582918829 . _:b582918830 . _:b582918831 . _:b582918824 . . _:b582918825 . _:b582918727 "3"^^ . . _:b582918826 . _:b582918827 . . _:b582918820 . _:b582918726 "3"^^ . _:b582918821 . _:b582918822 . _:b582918823 . . _:b42012404 . _:b582918816 . _:b582918721 "3"^^ . _:b582918917 . _:b582918817 . _:b582918818 . _:b582918819 . _:b582918844 . _:b582918667 . _:b582918916 . _:b582918845 . _:b582918720 "3"^^ . _:b582918846 . _:b582918847 . _:b582918840 . _:b582918935 . _:b582918723 "3"^^ . _:b582918919 . _:b582918841 . _:b582918842 . _:b582918843 . _:b582918836 . _:b582918722 "3"^^ . _:b582918918 . _:b582918837 . _:b582918838 . . _:b582918839 . _:b582918832 . _:b582918953 . _:b582918733 "3"^^ . _:b582918913 . _:b582918833 . _:b582918834 . _:b582918835 . _:b582918796 . _:b582918732 "3"^^ . _:b582918912 . _:b582918797 . _:b582918798 . _:b582918799 . _:b582918591 . _:b582918792 . _:b582918735 "3"^^ . _:b582918915 . _:b582918793 . _:b582918794 . _:b582918977 . _:b582918795 . _:b582918788 . _:b582918734 "3"^^ . _:b582918914 . _:b582918789 . _:b582918790 . _:b582918543 . _:b582918791 . . _:b582918784 . _:b582918729 "3"^^ . _:b582918925 . _:b582918785 . _:b582918786 . . _:b582918787 . _:b582918812 . . _:b582918924 . _:b582918813 . _:b582918728 "3"^^ . _:b582918611 . _:b582918814 . _:b582918815 . _:b582918808 . _:b582918731 "3"^^ . _:b582918927 . _:b582918809 . _:b582918810 . . _:b582918811 . _:b582918804 . _:b582918730 "3"^^ . _:b582918926 . _:b582918805 . . _:b582918806 . . _:b582918807 . _:b582918411 . _:b582918800 . _:b582918741 "3"^^ . _:b582919010 . _:b582918921 . . _:b582918801 . _:b582918802 . . _:b582918880 . _:b582918803 . _:b582918892 . _:b582918740 "3"^^ . _:b582918920 . _:b582918893 . _:b582918894 . . _:b582919030 . _:b582918895 . _:b582918888 . _:b582918743 "3"^^ . _:b582918923 . _:b582918889 . _:b582918890 . _:b582918476 . _:b582918891 . _:b582918884 . _:b582918742 "3"^^ . _:b582918922 . _:b582918885 . _:b582918886 . _:b582918887 . _:b582918880 . _:b582918737 "3"^^ . _:b582918933 . _:b582918881 . _:b582918882 . _:b582918479 . _:b582918883 . _:b582918626 . _:b582918908 . _:b582918736 "3"^^ . _:b582918932 . _:b582918909 . _:b582918910 . _:b582918803 . _:b582918614 . _:b582918911 . _:b582918904 . _:b582918739 "3"^^ . _:b582918935 . _:b582918905 . _:b582918448 . _:b582918906 . _:b582918907 . . _:b582918900 . _:b582918738 "3"^^ . _:b582918934 . _:b582918901 . . _:b582918902 . . . _:b582918903 . _:b582918814 . _:b582918896 . _:b582918749 "3"^^ . _:b582918929 . _:b582918897 . _:b582918898 . _:b582918899 . _:b582918860 . _:b582918748 "3"^^ . _:b582918928 . _:b582918861 . _:b582918862 . _:b582918989 . _:b582918863 . _:b582918856 . _:b582918751 "3"^^ . _:b582918931 . _:b582918857 . _:b582918858 . _:b582918859 . . _:b582918852 . _:b582918750 "3"^^ . _:b582918930 . _:b582918853 . _:b582918854 . _:b582919209 . _:b582918855 . _:b582918848 . _:b582918745 "3"^^ . _:b582918941 . _:b582918849 . . _:b582918850 . _:b582918851 . _:b582918876 . . _:b582918940 . _:b582918877 . . _:b582918744 "3"^^ . _:b582918878 . _:b582918879 . . . _:b582918872 . _:b582918747 "3"^^ . _:b582918943 . _:b582918873 . . _:b42012368 "As the protein stability of YAP/TAZ is regulated through phosphorylation by LATS1/2 kinases (Zhao et al. >>2010<<), we examined the phosphorylation status of endogenous YAP in TAZ(4SA)-overexpressing MCF10A cells." . _:b582918874 . _:b582918875 . _:b582918868 . _:b582918746 "3"^^ . _:b582918942 . _:b582918869 . _:b582918870 . _:b582918871 . . _:b582919218 . _:b582918864 . _:b582918757 "3"^^ . _:b582918937 . _:b582918865 . . _:b582918866 . _:b582918867 . _:b582918956 . _:b582918756 "3"^^ . _:b582918936 . _:b582918957 . _:b582918958 . _:b582918525 . _:b582918959 . _:b582918952 . _:b582918759 "3"^^ . _:b582918939 . _:b582918953 . _:b582918954 . _:b582918955 . _:b582918948 . _:b582918758 "3"^^ . _:b582918938 . _:b582918949 . _:b582918950 . _:b582918951 . _:b582918435 . _:b582918944 . _:b582918753 "3"^^ . _:b582918949 . _:b582918945 . _:b582918946 . _:b582918947 . _:b582918972 . _:b582918752 "3"^^ . _:b582918948 . _:b582918973 . _:b582918974 . . _:b582918975 . _:b582918968 . _:b582918755 "3"^^ . _:b582918951 . _:b582918969 . . _:b582918970 . _:b582918707 . _:b582918971 . _:b582918964 . _:b582918754 "3"^^ . _:b582918950 . _:b582918965 . _:b582918966 . _:b582918967 . _:b582918870 . _:b582918841 . _:b582918960 . _:b582918765 "3"^^ . _:b582918945 . _:b582918961 . _:b582918962 . _:b582918963 . _:b582918924 . _:b582919130 . . _:b582918944 . _:b582918925 . _:b582918764 "3"^^ . . _:b582918926 . _:b582918927 . _:b582918920 . _:b582918767 "3"^^ . _:b582918947 . _:b582918921 . . _:b582918922 . _:b582918923 . . _:b582918916 . _:b582918766 "3"^^ . _:b582918946 . _:b582918917 . _:b582918668 . _:b582918918 . _:b582918919 . _:b582918582 . _:b582918912 . _:b582918761 "3"^^ . _:b582918957 . _:b582918913 . _:b582918914 . _:b582918915 . _:b582918940 . . _:b582918760 "3"^^ . _:b582918956 . _:b582918898 . _:b582918941 . _:b582918942 . _:b582918943 . _:b582918936 . _:b582919204 . _:b582918763 "3"^^ . _:b582919016 . _:b582918959 . _:b582918937 . _:b582918697 . . _:b582918938 . . _:b582919201 . _:b582918872 . _:b582918939 . . _:b582918932 . _:b582918567 . . _:b582918958 . _:b582918604 . _:b582918933 . _:b582918762 "3"^^ . _:b582918934 . _:b582918935 . . _:b582918928 . _:b582918773 "3"^^ . _:b582918953 . _:b582918929 . _:b582918930 . _:b582918931 . _:b582919020 . _:b582918772 "3"^^ . _:b582918952 . _:b582919021 . _:b582919022 . _:b582919023 . _:b582918687 . _:b582919016 . _:b582918775 "3"^^ . _:b582918955 . _:b582919017 . _:b582919018 . . _:b582919019 . _:b582918556 . _:b582919012 . _:b582918774 "3"^^ . _:b582918954 . _:b582919013 . _:b582919014 . _:b582919015 . _:b582919008 . . _:b582918965 . _:b582919009 . _:b582918782 . _:b582918769 "3"^^ . _:b582919010 . . _:b582919011 . _:b582919036 . _:b582918768 "3"^^ . _:b582918964 . _:b582919037 . _:b582919038 . _:b582919039 . _:b582919032 . _:b582918771 "3"^^ . _:b582918967 . _:b582919033 . _:b582919034 . _:b582919035 . _:b582919028 . _:b582918770 "3"^^ . _:b582918966 . _:b582919029 . _:b582919030 . _:b582919031 . _:b582919024 . _:b582918781 "3"^^ . _:b582918961 . _:b582919025 . _:b582918457 . _:b582919026 . _:b582919027 . . _:b582918988 . _:b582918461 . _:b582918780 "3"^^ . _:b582918960 . _:b582918989 . _:b582918990 . . _:b582918990 . _:b582918991 . _:b582918685 . _:b42012401 . _:b582918984 . _:b582918783 "3"^^ . _:b582918963 . _:b582918985 . _:b582918986 . _:b42012384 . _:b582918987 . _:b582918980 . _:b582918782 "3"^^ . _:b582918962 . _:b582918981 . _:b582918982 . . _:b582918983 . _:b582918976 . _:b582919182 . _:b582918777 "3"^^ . _:b582918973 . _:b582918977 . _:b582918978 . _:b582918979 . . _:b582919004 . . _:b582918776 "3"^^ . _:b582918972 . _:b582919005 . _:b582919006 . _:b582919007 . _:b582919000 . _:b582918779 "3"^^ . _:b582918975 . _:b582919001 . _:b582918537 . _:b582919002 . _:b582919003 . . _:b582918996 . . _:b582918778 "3"^^ . _:b582918974 . _:b582918997 . _:b582918998 . _:b582918999 . _:b582918992 . _:b582918789 "3"^^ . _:b582918969 . _:b582918993 . _:b582918994 . _:b582918995 . _:b582919084 . _:b582918788 "3"^^ . _:b582918968 . _:b582919085 . _:b582919086 . _:b582919087 . _:b582919080 . _:b582918791 "3"^^ . _:b582918971 . _:b582919081 . _:b582919082 . _:b582919083 . _:b582919076 . _:b582919086 . _:b582918557 . _:b582918970 . _:b582919077 . _:b582918790 "3"^^ . _:b582919078 . . _:b582919079 . _:b582919072 . _:b582918785 "3"^^ . _:b582918981 . _:b582919073 . . _:b582919074 . _:b42012375 "Both YAP and TAZ can be potently dephosphorylated and activated by serum (Yu et al. >>2012<<). As expected, serum stimulation induced YAP dephosphorylation as determined by immunoblotting using an antibody specific to YAP phosphorylated at Ser127 (a direct LATS1/2 phosphorylation site responsible for cytoplasmic localization of" . . _:b582919075 . _:b582919100 . _:b582918809 . _:b582918980 . _:b582919101 . _:b582918784 "3"^^ . . . _:b582919102 . _:b582919103 . _:b582919096 . _:b582919049 . _:b582918787 "3"^^ . _:b42012382 "We then investigated whether this feedback mechanism operates in vivo using YAP transgenic (Tg) mice (Dong et al. >>2007<<). Consistent with what we observed with the cell lines in vitro, a significant increase of Lats2 mRNA, but not Lats1 mRNA, was detected in the livers of YAP Tg mice (Fig." . _:b582918983 . _:b582919097 . _:b582919098 . _:b582919099 . _:b582919092 . _:b582918786 "3"^^ . _:b582918982 . _:b582919093 . . _:b582919094 . _:b582919095 . _:b582918542 . _:b582919088 . _:b582918797 "3"^^ . _:b582918977 . _:b582918949 . _:b582919089 . _:b582918584 . _:b582919090 . . _:b582919091 . _:b582919052 . _:b582918796 "3"^^ . _:b582918976 . _:b582919053 . _:b582919054 . _:b582919055 . _:b582919048 . _:b582918799 "3"^^ . _:b582918979 . _:b582919049 . _:b582919050 . _:b582919051 . _:b582919044 . _:b582918798 "3"^^ . _:b582918978 . _:b582919045 . _:b582919046 . _:b582919047 . _:b582918633 . _:b582919040 . _:b582918793 "3"^^ . _:b582918989 . _:b582919041 . _:b582919042 . _:b582918503 . _:b582919043 . _:b582919068 . _:b582918792 "3"^^ . _:b582918988 . _:b582919069 . . _:b582919070 . . _:b582919071 . _:b582919064 . _:b582918795 "3"^^ . _:b582918991 . _:b582919065 . . _:b582919066 . . . _:b582919067 . . _:b582919060 . _:b582918794 "3"^^ . _:b582918990 . _:b582919061 . . _:b582919062 . _:b582919063 . _:b582919056 . . _:b582918985 . _:b582919057 . _:b582918805 "3"^^ . _:b582919058 . _:b582919059 . _:b582919148 . _:b582918804 "3"^^ . _:b582918984 . _:b582919149 . _:b582919150 . _:b582919151 . _:b582919144 . _:b582918807 "3"^^ . _:b582918987 . _:b582919145 . _:b582919146 . _:b582919147 . _:b582919140 . _:b42012363 "To achieve liver-specific or intestine-specific gene deletion, YapF/F mice were bred to Albumin-Cre (Zhang et al. 2010) or Villin-Cre (Taniguchi et al. >>2015<<) Tg mice, respectively. For liver-specific YAP Tg mice, 2-mo-old YAP Tg and non-Tg littermates were fed 0.2 mg/mL doxycycline (Sigma) in drinking water supplemented with 2.5% sucrose, as described previously (Dong et al. 2007)." . . _:b582918986 . _:b582919141 . _:b582918806 "3"^^ . _:b582919142 . . _:b582919143 . . _:b582919136 . . _:b582918801 "3"^^ . _:b582918997 . _:b582919137 . . _:b582919138 . _:b582919139 . _:b582918976 . _:b582919164 . _:b582918800 "3"^^ . _:b582918996 . _:b582919165 . _:b582919166 . _:b42012364 "For liver-specific YAP Tg mice, 2-mo-old YAP Tg and non-Tg littermates were fed 0.2 mg/mL doxycycline (Sigma) in drinking water supplemented with 2.5% sucrose, as described previously (Dong et al. >>2007<<). Two-month-old to 3-mo-old sex-matched mice were used for all experimental procedures." . _:b582919167 . . _:b582919160 . _:b582918803 "3"^^ . _:b582918999 . _:b582919161 . _:b582919162 . _:b582919163 . _:b582919156 . _:b582918802 "3"^^ . _:b582918998 . _:b582919157 . _:b582919158 . _:b582919159 . _:b582919203 . _:b582919152 . _:b582918813 "3"^^ . _:b582918993 . _:b582919153 . . _:b582919176 . _:b582919154 . _:b582919155 . _:b582919116 . _:b582918812 "3"^^ . _:b582918992 . _:b582919117 . _:b582919118 . _:b582919119 . _:b582919112 . _:b582919113 . _:b582918995 . _:b582918815 "3"^^ . _:b582919114 . _:b582919115 . _:b582919108 . _:b582919109 . _:b582918994 . _:b582918814 "3"^^ . _:b582919110 . _:b582918733 . . _:b582919111 . _:b582919104 . _:b582919105 . _:b582919005 . _:b582918809 "3"^^ . _:b582919106 . _:b582919123 . _:b582919107 . _:b582919132 . . _:b582919133 . _:b582919004 . _:b582918808 "3"^^ . _:b582919134 . _:b582919135 . _:b582919128 . _:b582919129 . _:b582919007 . _:b582918811 "3"^^ . _:b582919130 . _:b582919131 . _:b582919124 . _:b582919195 . _:b582919125 . _:b582919006 . . _:b582918810 "3"^^ . _:b582919126 . _:b582919127 . _:b582919120 . . _:b582919121 . _:b582919001 . _:b582918821 "3"^^ . . _:b582919122 . _:b582918836 . . _:b582919123 . . _:b582918717 . _:b582919142 . _:b582919000 . _:b582918820 "3"^^ . . _:b582918823 "3"^^ . _:b582919003 . _:b582918822 "3"^^ . _:b582919002 . _:b582918737 . _:b42012406 "In support of this idea, genetic mutations inhibiting the functions of key negative feedback regulators are found in human cancer: NF2 is mutated with high frequency in neurofibromatosis (Asthagiri et al. >>2009<<), and loss of NF2 as well as LATS2 is also frequently observed in malignant mesothelioma (Murakami et al. 2011), indicative of their functions as bona fide tumor suppressor genes." . . _:b582918817 "3"^^ . _:b582919013 . . _:b582918975 . . _:b582919012 . _:b582918816 "3"^^ . _:b582918887 . _:b582918654 . _:b582918819 "3"^^ . _:b582919015 . _:b582918859 . _:b582918818 "3"^^ . _:b582919014 . _:b582918820 . _:b582918829 "3"^^ . _:b42012374 . _:b582919009 . _:b582918828 "3"^^ . _:b582919008 . . _:b582918831 "2"^^ . . _:b582919011 . _:b582918605 . . . _:b582919010 . _:b582918830 "3"^^ . _:b42012399 "than other well-characterized negative feedback loops mediated by protein modifications, such as receptor phosphorylation by GPCR kinases (GRKs) followed by \u03B2-arrestin-mediated uncoupling of the GPCR from the G proteins (Pierce et al. >>2002<<) or feedback phosphorylation of upstream components of a signaling cascade (e.g., inhibitory phosphorylation of RAF by ERK in the MAPK signaling) (Dougherty et al. 2005)." . _:b582918689 . _:b582918825 "3"^^ . _:b42012370 . _:b582919021 . . _:b582918824 . _:b582918854 . _:b582918824 "3"^^ . _:b582919020 . _:b582918815 . _:b582919023 . _:b582918827 "3"^^ . _:b582918511 . _:b582918826 "3"^^ . _:b582919022 . . _:b582918837 "2"^^ . _:b582919017 . . . . _:b582918836 "2"^^ . _:b582919016 . _:b42012383 . _:b582918839 "2"^^ . _:b582919019 . _:b582918838 "2"^^ . _:b582918992 . _:b582919018 . _:b42012359 . . _:b582918833 "2"^^ . _:b582919029 . . _:b582918832 "2"^^ . _:b582919028 . _:b582918741 . _:b582918835 "2"^^ . _:b582918840 . _:b582919031 . . . _:b582918493 . _:b582918834 "2"^^ . _:b582919030 . . _:b582918504 . . _:b582918845 "2"^^ . _:b582919015 . _:b582919025 . _:b582918844 "2"^^ . _:b582919024 . _:b582918847 "2"^^ . _:b582919216 . _:b582919027 . _:b582919074 . _:b582918846 "2"^^ . _:b582919026 . _:b582918454 . _:b582918841 "2"^^ . . . _:b582919037 . _:b582918574 . _:b582918840 "2"^^ . . _:b582919036 . _:b582919019 . _:b582918843 "2"^^ . _:b582919039 . _:b582918842 "2"^^ . _:b582919038 . _:b42012379 . _:b582918853 "2"^^ . _:b582919033 . _:b582918756 . _:b582918852 "2"^^ . _:b582918890 . _:b582919032 . _:b582919207 . _:b582918855 "2"^^ . _:b582919035 . _:b582918936 . _:b582918854 "2"^^ . . _:b582919034 . _:b582918849 "2"^^ . _:b582919045 . . _:b582918848 "2"^^ . _:b582919044 . _:b582918682 . . _:b582918851 "2"^^ . _:b582919021 . _:b582919047 . _:b582919058 . . _:b582918850 "2"^^ . _:b582919046 . _:b582918861 "2"^^ . . _:b582919041 . _:b582918849 . . . _:b582918860 "2"^^ . _:b582919040 . _:b582918950 . _:b582918863 "2"^^ . _:b582919043 . _:b582918862 "2"^^ . _:b582919042 . . _:b582918857 "2"^^ . _:b582919053 . _:b582918856 "2"^^ . _:b582919052 . _:b582918859 "2"^^ . _:b582919055 . _:b582918858 "2"^^ . _:b582919054 . _:b582919112 . _:b582918869 "2"^^ . _:b582919049 . _:b582918868 "2"^^ . _:b582919039 . _:b582919048 . . _:b582918871 "2"^^ . . _:b582919051 . _:b582918870 "2"^^ . _:b582919050 . _:b582918865 "2"^^ . _:b582919061 . . _:b582918864 "2"^^ . _:b582919060 . _:b582918867 "2"^^ . _:b582919063 . . _:b582918608 . . _:b582918866 "2"^^ . _:b582919062 . _:b582918877 "2"^^ . . _:b582919057 . . . _:b582918710 . _:b582918876 "2"^^ . . _:b582919099 . _:b582919056 . _:b582918924 . . _:b582918879 "2"^^ . _:b582919059 . _:b582918638 . . _:b582918878 "2"^^ . _:b582919058 . _:b582918722 . _:b42012399 . . _:b582918873 "2"^^ . _:b582919069 . . . _:b582918872 "2"^^ . _:b582919068 . _:b582918651 . . . _:b582918875 "2"^^ . . _:b582919071 . _:b582919069 . . _:b582918874 "2"^^ . _:b582919070 . _:b582918885 "2"^^ . _:b582919065 . _:b582918444 . _:b582919197 . _:b582918884 "2"^^ . _:b582918445 . _:b582919064 . . _:b582918446 . _:b582918447 . _:b582919053 . _:b582918440 . _:b582918887 "2"^^ . _:b582918441 . _:b582919067 . . _:b582918442 . _:b582918443 . _:b582918436 . _:b582918886 "2"^^ . _:b582918437 . _:b582919066 . _:b582918438 . _:b582918439 . _:b582918432 . _:b582918881 "2"^^ . . _:b582918433 . _:b582919077 . _:b582918434 . _:b582918435 . _:b582918460 . _:b582918880 "2"^^ . _:b582918461 . _:b582919076 . _:b582918462 . _:b582918463 . _:b582918533 . _:b582918456 . _:b582918883 "2"^^ . . . _:b582918457 . _:b582919079 . _:b582918458 . _:b582919006 . _:b582918459 . . _:b582918452 . _:b582918882 "2"^^ . _:b582918453 . _:b582919078 . _:b582918454 . . _:b582918455 . _:b582918417 . _:b582918448 . _:b582918893 "2"^^ . _:b582918449 . _:b582919073 . _:b582918450 . . _:b582918451 . _:b582918412 . _:b582918892 "2"^^ . _:b582918875 . _:b582918413 . _:b582919072 . _:b582918414 . _:b582919060 . _:b582918415 . _:b582918895 "2"^^ . _:b582919075 . _:b582918410 . _:b582918411 . _:b582918894 "2"^^ . _:b582918983 . _:b582919074 . _:b582918889 "2"^^ . _:b582919085 . . _:b582918995 . _:b582918428 . _:b582918888 "2"^^ . _:b582918429 . _:b42012391 "studies revealed that the phosphorylation of AMOT by LATS1/2 inhibits AMOT binding to F-actin (Chan et al. 2013; Dai et al. 2013; Hirate et al. 2013), reduces actin stress fiber formation, and thereby inhibits migration (Dai et al. >>2013<<). We thus hypothesized that the LATS-mediated feedback mechanism limits cell migration induced by YAP activation." . _:b582919084 . _:b582918430 . . _:b582918431 . _:b582918424 . _:b582918891 "2"^^ . _:b582918425 . _:b582919087 . . _:b582918426 . . _:b582918427 . _:b582918420 . _:b582918890 "2"^^ . . . _:b582918421 . _:b582919086 . _:b582918422 . _:b582918423 . . _:b582918416 . _:b582918970 . _:b582918901 "2"^^ . _:b582918434 . _:b582919081 . _:b582918417 . _:b582918418 . . _:b582918419 . _:b582918508 . _:b582918900 "2"^^ . . _:b582918509 . _:b582919080 . _:b582918510 . _:b582918511 . _:b582918504 . _:b582918903 "2"^^ . _:b582918505 . _:b582919083 . _:b582918506 . _:b582918507 . _:b582918500 . _:b582918902 "2"^^ . _:b582918501 . _:b582919082 . . _:b582918786 . _:b582918502 . _:b582918503 . _:b582918496 . _:b582918897 "2"^^ . _:b582918497 . _:b582919093 . . _:b582918498 . _:b582918499 . _:b582918524 . _:b582919223 . _:b582918896 "2"^^ . _:b582918525 . _:b582919092 . _:b582918526 . _:b582918527 . _:b582918520 . _:b582918899 "2"^^ . _:b582918521 . _:b582919095 . _:b582918522 . _:b582918914 . _:b582918523 . _:b582918516 . _:b582918898 "2"^^ . _:b582918848 . _:b582918517 . _:b582919094 . _:b582918518 . _:b582918519 . _:b582918512 . _:b582918909 "2"^^ . _:b582918513 . _:b582919089 . _:b582918514 . _:b582918515 . _:b582918476 . _:b582918908 "2"^^ . _:b582918477 . _:b582919088 . _:b582918478 . _:b582918479 . . _:b582918472 . _:b582918911 "2"^^ . . _:b582918473 . _:b582919091 . _:b582918474 . _:b582918475 . _:b582918468 . _:b42012387 . _:b582918910 "2"^^ . . _:b582918469 . _:b582919090 . _:b582918470 . _:b42012401 "Notably, the increase of AMOT may also contribute to the negative feedback because AMOT can directly bind to and inhibit YAP/TAZ as well as indirectly activate LATS (Zhao et al. >>2011<<; Hirate et al. 2013)." . _:b582918471 . _:b582918464 . _:b582918905 "2"^^ . _:b582918465 . _:b582919101 . _:b582918466 . _:b582919133 . _:b582918833 . _:b582918571 . _:b582918467 . _:b582918492 . _:b582918904 "2"^^ . _:b582918590 . _:b582918493 . _:b582919100 . _:b582918494 . _:b582918495 . _:b582918915 . _:b582918488 . _:b582918907 "2"^^ . _:b582918489 . _:b582919103 . _:b582918490 . . _:b582918491 . _:b582918484 . _:b582918906 "2"^^ . _:b582918485 . _:b582919102 . _:b582918486 . _:b582918431 . _:b582918487 . _:b582918480 . _:b582918917 "2"^^ . . _:b582918481 . _:b582919097 . . _:b582918482 . _:b582918483 . _:b582919022 . _:b582918572 . _:b582918916 "2"^^ . _:b582918573 . _:b582919096 . _:b582918574 . _:b582918575 . _:b582918568 . _:b42012385 . _:b582918919 "2"^^ . _:b582918569 . _:b582919099 . _:b582918570 . _:b582918571 . _:b582918634 . _:b582918564 . _:b582918918 "2"^^ . . _:b42012377 "3F; Barretina et al. >>2012<<). As expected, there is a strong positive correlation between the two YAP/TAZ target genes CYR61 (cysteine-rich angiogenic inducer 61) and CTGF but no correlation between GAPDH (a negative control) and either CYR61 or CTGF." . _:b582919098 . _:b582918565 . _:b582918624 . _:b582918566 . _:b582918567 . _:b582918560 . _:b582918913 "2"^^ . _:b582918561 . _:b582919109 . _:b582918562 . _:b582918563 . _:b582918588 . _:b582918912 "2"^^ . _:b582918589 . _:b582919108 . _:b582918590 . _:b582918591 . _:b582918584 . _:b582918915 "2"^^ . . _:b582918585 . _:b582919111 . _:b582918586 . _:b582918587 . _:b582918580 . _:b42012393 . _:b582918914 "2"^^ . . _:b582918581 . _:b582919110 . _:b582918582 . _:b582918583 . _:b582918576 . _:b582918925 "2"^^ . _:b582918577 . _:b582919105 . _:b582918578 . _:b582918579 . _:b582918540 . _:b582918924 "2"^^ . _:b582918541 . _:b582919104 . _:b582918542 . _:b582918543 . _:b582919222 . _:b582918932 . _:b582918536 . _:b582918927 "2"^^ . _:b582918871 . . _:b582918537 . _:b582919107 . _:b582918538 . . _:b582918539 . _:b582918698 . _:b582918532 . _:b582918926 "2"^^ . _:b582918533 . _:b582919106 . _:b582918785 . _:b582918534 . _:b582918535 . _:b582918528 . _:b42012375 . _:b582918921 "2"^^ . _:b582918529 . _:b582919117 . _:b582918530 . _:b582918531 . _:b582918556 . _:b582918920 "2"^^ . _:b582918557 . _:b582919116 . _:b582918558 . _:b582918559 . _:b582919164 . _:b582918552 . _:b582918923 "2"^^ . _:b582918553 . _:b582919119 . _:b582918554 . _:b582918555 . _:b582918858 . _:b582918548 . _:b582918922 "2"^^ . _:b582918463 . . _:b582919118 . _:b582918549 . _:b582918550 . _:b582918793 . _:b582918711 . _:b582918551 . _:b582918544 . _:b582918933 "2"^^ . _:b582918545 . _:b582919113 . _:b582918546 . _:b582918538 . _:b582918547 . _:b582918636 . _:b582918932 "2"^^ . _:b582918637 . _:b582919112 . . _:b582919104 . _:b582918638 . _:b582918639 . _:b582918632 . _:b582918935 "2"^^ . _:b582918633 . _:b582919115 . _:b582918634 . _:b582918635 . . _:b582918934 "2"^^ . _:b582918628 . _:b582918629 . _:b582919114 . _:b582918630 . _:b582918570 . _:b582918631 . _:b582919206 . _:b582918624 . _:b582919171 . _:b582918929 "2"^^ . _:b582918625 . _:b582919125 . _:b582918626 . _:b582918627 . _:b582918652 . _:b582918928 "2"^^ . _:b582918653 . _:b582919124 . _:b582918654 . _:b582919029 . . _:b582918655 . _:b582918648 . _:b582918931 "2"^^ . . _:b582918778 . _:b582918649 . _:b582919127 . . _:b582918650 . _:b582918651 . _:b582918569 . _:b582918644 . _:b582918930 "2"^^ . _:b582918645 . _:b582919126 . _:b582918646 . _:b42012394 "feedback mechanism employed by the mammalian Hippo pathway appears to differ significantly from that in D. melanogaster, where Yorkie induces expression of expanded, merlin, four-jointed, or kibra (Cho et al. 2006; Hamaratoglu et al. >>2006<<; Genevet et al. 2010), but not warts (the fly homolog of LATS1/2), in dividing epithelial cells of the wing or eye disc (Jukam et al. 2013)." . _:b582918647 . _:b582918640 . _:b582918941 "2"^^ . _:b582918641 . _:b582919121 . _:b582918642 . . _:b582918643 . _:b582918604 . _:b582918940 "2"^^ . . _:b582918605 . _:b582919120 . _:b582918606 . . _:b582918607 . _:b582918600 . _:b582919121 . _:b582918943 "2"^^ . _:b582918601 . _:b582919123 . _:b582918602 . _:b582918603 . _:b582918447 . _:b582918596 . _:b582918942 "2"^^ . _:b582918597 . _:b582919122 . _:b582918598 . _:b582918599 . _:b582918592 . _:b582918937 "2"^^ . . _:b582919133 . _:b582918593 . _:b582918594 . _:b582918595 . . _:b582918620 . _:b582918936 "2"^^ . _:b582918621 . _:b582919132 . _:b582918622 . _:b582918623 . _:b582918616 . _:b582918939 "2"^^ . _:b582918617 . _:b582919135 . _:b582918618 . _:b582918619 . _:b582918612 . _:b582918938 "2"^^ . _:b582918613 . _:b42012400 . _:b582919134 . _:b582918614 . _:b582918615 . _:b582918608 . _:b582918949 "2"^^ . _:b582918609 . _:b582919129 . _:b582918610 . _:b582918611 . _:b582918948 "2"^^ . . _:b582919128 . _:b582918652 . _:b582918579 . _:b582918951 "2"^^ . _:b582919131 . _:b582918950 "2"^^ . _:b582919130 . _:b582918940 . _:b582918945 "2"^^ . . _:b582919141 . _:b582918623 . _:b582918643 . _:b582918944 "2"^^ . _:b582919140 . _:b582918947 "2"^^ . _:b582918997 . _:b582919143 . _:b582918946 "2"^^ . _:b582919142 . _:b582918957 "2"^^ . _:b582919137 . _:b582918956 "2"^^ . _:b582919136 . . _:b582918959 "2"^^ . _:b582919077 . _:b582919139 . _:b582918958 "2"^^ . _:b582919138 . _:b582918953 "2"^^ . _:b582919149 . _:b582918719 . . _:b582919051 . _:b582918952 "2"^^ . _:b582919148 . _:b42012378 . _:b582918955 "2"^^ . _:b42012386 . _:b582919151 . . _:b582918954 "2"^^ . _:b582919150 . _:b582918965 "2"^^ . _:b582919070 . _:b582919145 . _:b582918964 "2"^^ . _:b582919144 . . . _:b582919155 . _:b582918967 "2"^^ . . _:b582919147 . . _:b582918966 "2"^^ . _:b582919146 . _:b582918774 . . . _:b582918961 "2"^^ . _:b582919157 . _:b582918945 . _:b582918960 "2"^^ . _:b582919156 . _:b582919013 . _:b582918963 "2"^^ . _:b582919159 . . _:b582918986 . . _:b582918962 "2"^^ . . . _:b582919158 . _:b582918973 "2"^^ . _:b582919153 . . _:b582918972 "2"^^ . . _:b582919152 . _:b582918650 . _:b582918975 "2"^^ . _:b582919155 . _:b582918974 "2"^^ . _:b582919154 . . . . _:b582918600 . _:b582918969 "2"^^ . _:b582919165 . _:b582918968 "2"^^ . _:b582919164 . . . . _:b582918971 "2"^^ . . . _:b582919167 . . _:b582918970 "2"^^ . . _:b582919166 . . _:b582918981 "2"^^ . _:b582919161 . _:b582919046 . _:b582918980 "2"^^ . _:b582919160 . _:b582918951 . . _:b582918983 "2"^^ . _:b582919163 . . _:b582918982 "2"^^ . _:b582919162 . _:b582918977 "2"^^ . _:b582918661 . _:b582918976 "2"^^ . _:b582918660 . . _:b582918979 "2"^^ . . _:b582918663 . . _:b582918978 "2"^^ . . _:b582918662 . _:b582918989 "2"^^ . _:b582918657 . . _:b582918988 "2"^^ . _:b582918656 . . _:b582918991 "2"^^ . _:b582918637 . _:b582918659 . . _:b582918990 "2"^^ . _:b582918658 . _:b582918985 "2"^^ . . _:b582918669 . . _:b582919040 . _:b582918984 "2"^^ . _:b582918668 . _:b582918987 "2"^^ . _:b582918671 . . _:b582918986 "2"^^ . _:b582918670 . _:b582918560 . . _:b582918997 "2"^^ . _:b582918665 . _:b582918649 . _:b42012381 "Furthermore, Merlin is already an established transcriptional target of Yorkie, the fly homolog of YAP/TAZ (Hamaratoglu et al. >>2006<<). Indeed, we found that mRNA and protein abundance of NF2 both were markedly increased by YAP(5SA) or TAZ(4SA) overexpression in MCF10A cells (Fig. 4C,D; Supplemental Fig. S4D). ChIP assay revealed that both endogenous YAP and TEAD1" . . _:b582918996 "2"^^ . . _:b582918721 . _:b582918664 . . _:b582918999 "2"^^ . _:b582918667 . _:b582918607 . _:b582918998 "2"^^ . _:b582918666 . . _:b582918993 "2"^^ . _:b582918677 . . _:b582918992 "2"^^ . _:b582918676 . _:b582918995 "2"^^ . . _:b582918679 . . _:b582918994 "2"^^ . _:b582918678 . _:b582918808 . . _:b582919005 "2"^^ . . _:b582918673 . _:b582918879 . _:b582918764 . _:b582918844 . _:b582919004 "2"^^ . _:b582918672 . _:b582918906 . _:b582919007 "2"^^ . _:b582918675 . _:b582918881 . _:b582919006 "2"^^ . . _:b582918674 . . _:b582919001 "2"^^ . _:b582918685 . _:b582919000 "2"^^ . _:b582918684 . _:b582919003 "2"^^ . _:b582918687 . _:b582918535 . _:b582919002 "2"^^ . _:b582918686 . _:b42012369 . _:b582919013 "2"^^ . _:b582918681 . . . _:b582919012 "2"^^ . _:b582918680 . _:b582919015 "2"^^ . _:b42012363 . _:b582918683 . _:b582918846 . _:b582919014 "2"^^ . _:b582918682 . _:b582919009 "2"^^ . . _:b582918693 . _:b582919140 . _:b582918909 . _:b582919008 "2"^^ . _:b582918692 . . _:b582919011 "2"^^ . _:b582918695 . . _:b582919010 "2"^^ . _:b582918694 . . _:b582919021 "2"^^ . _:b582918689 . _:b582919020 "2"^^ . . _:b582918688 . . _:b582919023 "2"^^ . _:b582918691 . _:b582919022 "2"^^ . _:b582918690 . . _:b582919017 "2"^^ . _:b582918701 . . _:b582919016 "2"^^ . _:b582918700 . _:b582919019 "2"^^ . _:b582918514 . _:b582918703 . . . _:b582919018 "2"^^ . _:b582918702 . . _:b582919029 "2"^^ . . _:b582918697 . . . _:b582919028 "2"^^ . _:b582918696 . . _:b582919031 "2"^^ . _:b582918699 . _:b582919030 "2"^^ . _:b582918698 . _:b582918823 . _:b582918886 . _:b582919025 "2"^^ . _:b582918709 . . _:b582919024 "2"^^ . _:b42012359 "LATS1/2-, MST1/2-, or NF2-deficient HEK293A cells were created through the CRISPR/Cas9 system (Ran et al. >>2013<<). HEK293A cells were transfected with a Cas9 expression plasmid together with single-guide RNA (sgRNA) expression plasmids (PX459; Addgene plasmid no." . . _:b582918708 . _:b582918631 . . _:b582919027 "2"^^ . _:b582918711 . . _:b582919026 "2"^^ . _:b582918710 . . . _:b582919037 "2"^^ . _:b582918705 . . _:b582919036 "2"^^ . _:b582918704 . _:b582919039 "2"^^ . _:b582918707 . _:b582919115 . _:b582919038 "2"^^ . _:b582918706 . _:b582919141 . _:b582919110 . _:b582919033 "2"^^ . . _:b582918717 . _:b582918991 . _:b582919032 "2"^^ . _:b582918716 . _:b582918714 . . _:b582918512 . _:b582919035 "2"^^ . . _:b582918719 . . . _:b582919034 "2"^^ . _:b582918718 . _:b582919037 . _:b582919045 "2"^^ . _:b582918498 . _:b582918713 . _:b582919044 "2"^^ . . _:b582918712 . . _:b582919047 "2"^^ . _:b582918715 . _:b582919234 . . _:b582919226 . _:b582919066 . _:b582919046 "2"^^ . _:b582918714 . _:b582919041 "2"^^ . . _:b582918725 . _:b582919040 "2"^^ . . _:b582918724 . . _:b582919043 "2"^^ . . _:b582918727 . _:b582918954 . _:b582919042 "2"^^ . _:b582918726 . _:b582918440 . _:b582918740 . _:b582919053 "2"^^ . . _:b582918721 . _:b582918544 . _:b582918943 . _:b582919052 "2"^^ . _:b582918720 . _:b582919065 . . _:b582918752 . _:b582918957 . _:b582919055 "2"^^ . _:b582918723 . _:b582918787 . _:b582919041 . _:b582919054 "2"^^ . _:b582918722 . _:b582918443 . . _:b582919049 "2"^^ . _:b582918733 . . _:b582918925 . _:b582919048 "2"^^ . _:b582918732 . . _:b582919051 "2"^^ . _:b582918735 . . _:b582919050 "2"^^ . _:b582918734 . _:b582919061 "2"^^ . _:b582918729 . _:b582918755 . . _:b582919228 . _:b582919060 "2"^^ . _:b582918728 . . . _:b582919063 "2"^^ . _:b582918731 . _:b582919062 "2"^^ . _:b582918730 . _:b582919057 "2"^^ . _:b582918741 . _:b582918882 . _:b582919056 "2"^^ . _:b582918740 . . . _:b582919059 "2"^^ . _:b582918743 . . _:b582919058 "2"^^ . _:b582918742 . . _:b582918912 . _:b582919069 "2"^^ . _:b582918737 . _:b582918918 . _:b582919068 "2"^^ . _:b582918736 . _:b582918715 . . _:b582919071 "2"^^ . _:b582918739 . . _:b582919159 . _:b582919070 "2"^^ . _:b582918738 . _:b582919065 "2"^^ . _:b582918749 . . _:b582918939 . _:b582918426 . _:b582919064 "2"^^ . _:b582918748 . _:b582918758 . _:b582918665 . _:b582919122 . _:b582919067 "2"^^ . _:b582918751 . . _:b582919066 "2"^^ . _:b582918750 . _:b582919009 . _:b582919077 "2"^^ . _:b582918745 . _:b582919076 "2"^^ . _:b582918744 . _:b582918994 . _:b42012406 . _:b582919079 "2"^^ . _:b582918747 . _:b582918593 . _:b582918894 . _:b582919078 "2"^^ . _:b582918746 . . _:b582918795 . _:b582919073 "2"^^ . _:b582918757 . _:b582919072 "2"^^ . _:b582918756 . _:b42012405 . _:b582919075 "2"^^ . _:b582918759 . . _:b582918653 . _:b582919074 "2"^^ . _:b582918758 . _:b582918494 . _:b582919085 "2"^^ . _:b582918753 . _:b582919084 "2"^^ . _:b582918752 . . _:b582919087 "2"^^ . _:b582918755 . . _:b582919086 "2"^^ . _:b582918754 . _:b582919103 . _:b582919081 "2"^^ . . _:b582918765 . _:b582919080 "2"^^ . _:b582918764 . . _:b582918922 . _:b582919083 "2"^^ . _:b582918767 . _:b582919082 "2"^^ . _:b582918766 . _:b582918958 . _:b582919050 . _:b582919093 "2"^^ . _:b582918761 . . . _:b582919092 "2"^^ . _:b582918760 . _:b582918490 . . _:b582919095 "2"^^ . _:b582918763 . _:b582918873 . _:b582919094 "2"^^ . _:b582918762 . _:b42012358 "materials and methods" . _:b582918460 . _:b582919089 "2"^^ . _:b582918773 . . _:b582919088 "2"^^ . _:b582918772 . _:b582918636 . _:b582919091 "2"^^ . _:b582918775 . _:b582919090 "2"^^ . _:b582918774 . . _:b582919101 "2"^^ . _:b582918769 . _:b582918996 . . _:b582919100 "2"^^ . _:b582919127 . _:b582918768 . . _:b582918518 . _:b582919103 "2"^^ . _:b582918771 . . . _:b582919102 "2"^^ . _:b582918770 . _:b582919113 . . _:b582919097 "2"^^ . _:b582918781 . _:b582918828 . _:b582919096 "2"^^ . _:b582918780 . _:b582918908 . . _:b582919099 "2"^^ . _:b582918783 . _:b582918934 . . . _:b582919098 "2"^^ . _:b582918782 . _:b582918830 . _:b582919109 "2"^^ . _:b582918777 . _:b582919108 "2"^^ . _:b582918776 . _:b582919111 "2"^^ . _:b582918779 . _:b582918998 . _:b42012376 "CTGF (connective tissue growth factor), a known direct target gene of the YAP\u2013TEAD complex (Zhao et al. >>2008<<), and GAPDH (glyceraldehyde 3-phosphate dehydrogenase) were used as a positive and negative control, respectively." . _:b582919110 "2"^^ . _:b582918778 . _:b582918720 . . _:b582919105 "2"^^ . _:b582918789 . _:b582918716 . _:b582919104 "2"^^ . _:b582918788 . _:b582919107 "2"^^ . _:b582918791 . _:b582918952 . _:b582919106 "2"^^ . _:b582918790 . _:b582918548 . _:b582918597 . . _:b582919117 "2"^^ . _:b582918785 . . _:b582918920 . _:b582919116 "2"^^ . _:b582918784 . _:b582918966 . _:b582919119 "2"^^ . _:b582918787 . . _:b582919118 "2"^^ . _:b582918786 . . _:b582919113 "2"^^ . _:b582918797 . . _:b582919112 "2"^^ . _:b582918796 . _:b582919115 "2"^^ . _:b582918799 . . . _:b582919114 "2"^^ . _:b582918798 . . _:b582918684 . _:b582918606 . _:b582919125 "2"^^ . _:b582918793 . _:b582919124 "2"^^ . _:b582918792 . _:b582918673 . . _:b582918478 . _:b582919127 "2"^^ . _:b582918795 . _:b582919126 "2"^^ . _:b582918794 . _:b582918706 . . _:b582919121 "2"^^ . _:b582918805 . _:b582918449 . _:b582918712 . _:b582919055 . _:b582919120 "2"^^ . _:b582918804 . _:b582919094 . _:b582918754 . _:b582918974 . _:b582919123 "2"^^ . _:b582918807 . _:b582919122 "2"^^ . _:b582918806 . _:b582918522 . _:b582919133 "2"^^ . _:b582918801 . _:b582919002 . . _:b582919132 "2"^^ . _:b582918800 . _:b582918647 . _:b582918495 . _:b582919135 "2"^^ . _:b582918803 . _:b582919134 "2"^^ . _:b582918802 . _:b582919129 "2"^^ . _:b582918813 . _:b582919025 . _:b582919128 "2"^^ . _:b582918812 . _:b582918555 . _:b582919131 "2"^^ . _:b582918815 . . _:b582919130 "2"^^ . _:b582918814 . . _:b582918536 . _:b42012405 "YAP/TAZ activation in human cancers, the general lack of cancer-relevant somatic or germline mutations in core members of the Hippo pathway is a major conundrum to the field (Harvey et al. 2013; Moroishi et al. 2015; Plouffe et al. >>2015<<). One possibility is that negative feedback control of YAP/TAZ activity suppresses tumorigenesis following small perturbation of the pathway." . _:b582919141 "2"^^ . _:b582918809 . _:b582919140 "2"^^ . _:b582918808 . _:b582919198 . _:b582919143 "2"^^ . _:b582918811 . _:b582919081 . _:b582918759 . _:b42012397 "On the contrary, Yorkie indirectly suppresses Warts transcription to establish positive feedback regulation in combination with tissue-specific transcription factors in the context of Drosophila eye development (Jukam et al. >>2013<<). This is in a dramatic contrast to mammalian cells in which YAP/TAZ activate LATS. It is possible that the mechanism observed in D. melanogaster might be employed in some mammalian cell types. Although the evolutionary conservation of" . _:b582919188 . _:b582919142 "2"^^ . _:b582918810 . . _:b582919137 "2"^^ . _:b582918821 . _:b582919136 "2"^^ . _:b582918820 . . _:b582919035 . . _:b582919139 "2"^^ . _:b582918724 . _:b582918823 . _:b582918749 . _:b582919138 "2"^^ . _:b582918822 . . _:b582919149 "2"^^ . _:b582918817 . _:b582919148 "2"^^ . _:b582918816 . . . _:b582919151 "2"^^ . _:b582918819 . _:b582919150 "2"^^ . _:b582918818 . _:b582919145 "2"^^ . . _:b582918829 . _:b582919144 "2"^^ . . _:b582918828 . _:b582919147 "2"^^ . _:b582918831 . _:b582919146 "2"^^ . _:b582918830 . _:b582918751 . . _:b582918961 . _:b582919157 "2"^^ . . _:b582918825 . _:b582918580 . _:b582919156 "2"^^ . . _:b582918824 . _:b582919159 "2"^^ . _:b582918827 . . _:b582919158 "2"^^ . _:b582918826 . . _:b582919153 "2"^^ . . _:b582918837 . _:b582919059 . _:b582918413 . _:b582919152 "2"^^ . _:b582918836 . _:b582918470 . _:b582919155 "2"^^ . _:b582918839 . . _:b582919154 "2"^^ . _:b582918838 . _:b582919092 . _:b582919165 "2"^^ . _:b582918572 . _:b582918833 . _:b42012383 "Notably, the protein abundance of AMOT was also elevated in the YAP Tg liver, consistent with the previous observation that the phosphorylation stabilizes AMOT (Adler et al. >>2013<<). Collectively, these data support a model in which YAP/TAZ induce LATS activation to constitute a feedback mechanism in the Hippo pathway." . _:b582919164 "2"^^ . _:b582918832 . _:b582919167 "2"^^ . _:b582918835 . _:b582919166 "2"^^ . _:b582918834 . . _:b582919161 "2"^^ . _:b582918845 . _:b582919160 "2"^^ . . _:b582918844 . _:b582919163 "2"^^ . _:b582918847 . _:b582919162 "2"^^ . _:b42012403 . _:b582918846 . _:b582918841 . . . _:b582918840 . _:b582919167 . _:b42012386 "AMOT is a filamentous actin (F-actin)-binding protein. While YAP and TAZ are known to promote cell migration (Chan et al. >>2008<<; Haskins et al. 2014), recent studies revealed that the phosphorylation of AMOT by LATS1/2 inhibits AMOT binding to F-actin (Chan et al. 2013; Dai et al. 2013; Hirate et al. 2013), reduces actin stress fiber formation, and thereby" . _:b582918843 . . _:b582918842 . _:b582918853 . _:b582918897 . _:b582918852 . . _:b582918903 . _:b582918855 . _:b582918973 . . _:b582918854 . _:b582918855 . _:b582918541 . _:b582918849 . . . _:b582918452 . _:b582918888 . _:b582918848 . _:b582918547 . . _:b582918962 . _:b582918851 . . _:b582918666 . _:b582918850 . . _:b582918780 . . _:b582918861 . . . . _:b582918876 . _:b582918860 . _:b582918863 . . _:b582918806 . . _:b582918862 . . _:b582918857 . . _:b582918856 . . _:b582918859 . _:b582918858 . _:b582919162 . _:b582918418 . _:b582918869 . . _:b582918868 . _:b582918686 . _:b582918871 . _:b42012365 "results" . _:b582918870 . _:b582918700 . _:b582919116 . . . _:b582918865 . _:b582918410 . _:b582918866 . _:b582918864 . _:b582918867 . _:b582918866 . _:b582918669 . _:b582918681 . _:b582918877 . _:b582918876 . . _:b582918879 . _:b582919233 . _:b582918878 . _:b42012361 . _:b582918873 . . _:b582918872 . _:b42012398 "This evolutionary divergence in pathway components adds an extra layer of complexity and contributes to the dynamic regulation of the mammalian Hippo pathway (Bossuyt et al. >>2014<<)." . . . _:b582918875 . . _:b582919085 . _:b582918874 . _:b582918438 . _:b582918851 . . _:b582918885 . _:b582919174 . . _:b582918884 . _:b582918916 . . _:b582918887 . _:b582918621 . _:b582918886 . _:b582918881 . . _:b582918880 . . _:b582918822 . . _:b582919108 . _:b582918883 . _:b582918475 . _:b582918412 . _:b582918693 . _:b582918882 . _:b582919212 . _:b582919034 . . _:b582919100 . _:b582918893 . . . _:b582918892 . . . . _:b582918895 . . _:b582918894 . . . _:b582918889 . . _:b582918888 . _:b582919183 . _:b582918891 . _:b582918890 . _:b582918901 . . _:b582918900 . _:b582918789 . . _:b582918903 . . _:b582918902 . _:b582918897 . _:b582919125 . . . _:b582918812 . _:b582918896 . . _:b582918899 . . . _:b582918898 . _:b582918734 . _:b582918909 . _:b582919232 . _:b582918908 . _:b582919170 . _:b582918911 . . _:b582918910 . _:b582918765 . _:b582918905 . . _:b582918904 . _:b582918598 . _:b582918907 . . _:b582918906 . _:b42012392 . . . . . _:b582918573 . _:b582919178 . . . _:b42012365 . _:b582918413 . _:b582918526 . _:b582918412 . . _:b42012373 "Consistent with the observed LATS1 and LATS2 activation, phosphorylation of angiomotin (AMOT) at Ser175, a known direct substrate site for LATS1/2 kinases (Adler et al. 2013; Chan et al. 2013; Dai et al. 2013; Hirate et al. >>2013<<), was increased in TAZ(4SA)-overexpressing MCF10A cells (Fig. 2E). Moreover, the protein abundance of AMOT was also increased without a corresponding effect on the amount of AMOT mRNA (Supplemental Fig." . _:b42012358 . _:b582918415 . _:b582918414 . . . _:b582918968 . _:b582919023 . _:b582918552 . . _:b582918723 . _:b582918971 . _:b582918411 . _:b582918410 . . _:b582918421 . . . _:b582918420 . . _:b582918423 . _:b582919052 . _:b582918655 . _:b582918619 . _:b582918628 . _:b582919090 . _:b582918422 . _:b582918891 . _:b582918742 . . _:b42012368 . _:b582918417 . _:b582918416 . . _:b582918674 . _:b582918834 . _:b582918419 . . _:b42012390 "TAZ are known to promote cell migration (Chan et al. 2008; Haskins et al. 2014), recent studies revealed that the phosphorylation of AMOT by LATS1/2 inhibits AMOT binding to F-actin (Chan et al. 2013; Dai et al. 2013; Hirate et al. >>2013<<), reduces actin stress fiber formation, and thereby inhibits migration (Dai et al. 2013)." . _:b582918418 . _:b582918429 . . _:b582918428 . _:b42012407 . . _:b582919131 . _:b582918431 . . . _:b582918430 . . _:b582918982 . _:b582918425 . . . . _:b582918424 . . _:b582918837 . _:b582918427 . _:b582918519 . _:b582918426 . _:b582919220 . _:b582918437 . _:b582918777 . . _:b582918444 . . _:b582919230 . _:b582918436 . _:b582918853 . . _:b582918439 . _:b582918577 . . _:b582918618 . _:b582918438 . _:b582918433 . _:b42012362 . _:b582918432 . . _:b582918726 . . _:b582918435 . _:b582918434 . . _:b582918445 . _:b582918985 . _:b582918444 . _:b582918501 . . . . _:b582918447 . _:b582919156 . _:b582918446 . _:b582918499 . _:b582918441 . _:b582918440 . . _:b582918443 . _:b582919208 . _:b582918442 . _:b42012360 . _:b582918453 . _:b582918492 . _:b582918452 . _:b582919148 . _:b582918857 . _:b582918455 . _:b582918690 . _:b582918832 . _:b582918454 . . _:b582918449 . _:b582918448 . _:b582918955 . _:b582919026 . _:b582918451 . _:b582918450 . . _:b582918461 . _:b582919089 . _:b582918460 . _:b582919118 . _:b582918463 . _:b582918691 . _:b582918587 . _:b582918852 . _:b582918462 . . . _:b582918632 . _:b582918451 . _:b582918457 . . . _:b582918456 . . _:b582918804 . _:b582918459 . . _:b582918458 . _:b582918520 . _:b582918672 . _:b582918794 . _:b582918469 . . _:b582918468 . . . _:b582918592 . . _:b582918471 . _:b582918470 . _:b42012360 "org/crispr or were taken from archived guide sequences from the genome-scale CRISPR knockout (GeCKO2) library (Sanjana et al. >>2014<<). The guide sequences used were 5\u2032-CGTGCAGCTCTCCGCTCTAA-3\u2032 for human LATS1, 5\u2032-TACGCTGGCACCGTAGCCCT-3\u2032 for human LATS2, 5\u2032-ATACACCGAGATATCAAGGC-3\u2032 for human MST1, 5\u2032-AGTACTCCATAACAATCCAG-3\u2032 for human MST2, and 5\u2032-GTCCATGGTGACGATCCTCA-3\u2032" . _:b582918465 . _:b582918524 . _:b582918662 . _:b582918464 . _:b582918506 . _:b582918467 . _:b582919126 . _:b582918466 . . . _:b582918781 . _:b582918477 . . _:b582919135 . _:b582918476 . _:b582918479 . _:b582918728 . _:b582919175 . . _:b582918816 . _:b582918478 . . _:b582918473 . . _:b582919084 . . _:b582918472 . _:b582918864 . _:b582918475 . _:b582918792 . _:b582918485 . . _:b582918474 . _:b582919154 . _:b582918485 . _:b582918583 . _:b582918433 . _:b582918531 . _:b582918609 . _:b582918484 . . _:b582918487 . _:b582918486 . _:b582918481 . . _:b582918480 . . . _:b582918483 . _:b582918868 . _:b582918482 . _:b582918746 . _:b582918867 . _:b582918493 . _:b582919120 . . _:b42012379 "To test this possibility, we deleted MST1/2 in HEK293A cells using CRISPR/Cas9 genome-editing technology (Ran et al. >>2013<<). Consistent with a positive role for MST1/2 in YAP phosphorylation, MST1/2 deletion reduced the basal phosphorylation of YAP (Supplemental Fig. S4A). To avoid the influences from off-target effects, we obtained multiple clones and" . . _:b582918702 . _:b582918492 . _:b582918495 . _:b582918967 . _:b582919045 . _:b582918494 . _:b582918446 . . . _:b582919096 . _:b582918489 . _:b42012367 "a codependent regulatory relationship between Yorkie (the fly homolog of YAP/TAZ) and dMyc (encoded by the diminutive gene) in which Yorkie induces dMyc transcription, which in turn represses Yorkie expression (Neto-Silva et al. >>2010<<). However, the mRNA abundance of MYC was not increased, but rather decreased, in YAP(5SA)- or TAZ(4SA)-overexpressing MCF10A cells (Supplemental Fig." . _:b582919172 . _:b582918488 . . _:b582918491 . . _:b582918490 . _:b582918501 . _:b582919083 . _:b582918462 . _:b582918496 . . _:b582918500 . . _:b582918503 . _:b582918502 . _:b582918497 . . . . _:b582918496 . . _:b582918499 . . _:b42012390 . _:b582918498 . _:b582918509 . _:b582918508 . _:b582919163 . _:b582918511 . _:b42012381 . _:b582918510 . _:b582918505 . . _:b582918826 . _:b582918504 . . _:b582918507 . . . _:b582918506 . _:b582918517 . . _:b582919211 . . _:b582918516 . _:b582918519 . _:b582918518 . _:b42012402 . _:b582918513 . . _:b582918512 . _:b582918515 . . _:b582918515 . _:b582918514 . . _:b582918525 . . . _:b582918524 . _:b42012394 . _:b582919157 . _:b582918489 . . _:b582918527 . . _:b582918526 . . _:b582918521 . . . . _:b582918520 . _:b42012395 . . _:b582918523 . _:b582918602 . . _:b582918522 . _:b582919012 . _:b582918533 . . _:b42012377 . _:b582918532 . _:b582918535 . _:b582918988 . . . _:b582918534 . . _:b582918529 . _:b582918972 . _:b582918469 . . _:b582918528 . _:b582918827 . _:b582918531 . _:b582919214 . _:b582918578 . _:b582918530 . _:b582918486 . . _:b582918541 . _:b582918540 . . _:b582918543 . _:b582918471 . . _:b582918546 . _:b582918542 . _:b582918537 . _:b582919011 . _:b582918896 . _:b582918536 . _:b582918539 . _:b582918428 . _:b582919054 . _:b582918538 . _:b582918549 . . _:b582919000 . . _:b582918548 . _:b582918551 . _:b582918763 . _:b582918550 . . _:b582918878 . _:b582918798 . _:b582918545 . . . _:b582918544 . _:b582918612 . _:b582918547 . . . _:b582919173 "2"^^ . _:b582918546 . _:b582918557 . _:b42012373 . _:b582919172 "2"^^ . _:b582919024 . _:b582919088 . . _:b582918556 . _:b582919175 "2"^^ . . _:b582918559 . _:b582919174 "2"^^ . _:b582918944 . . _:b582918558 . _:b582919169 "2"^^ . _:b582918553 . _:b582919168 "2"^^ . _:b582918656 . . _:b582918552 . _:b582919171 "2"^^ . _:b582918555 . _:b582919170 "2"^^ . _:b582918497 . _:b582918554 . _:b582919181 "2"^^ . . . _:b582918565 . _:b582919180 "2"^^ . _:b582918564 . _:b582919229 . _:b582919183 "2"^^ . _:b582918567 . _:b582919182 "2"^^ . _:b582919014 . . _:b582918566 . _:b582919177 "2"^^ . _:b582918561 . _:b582919176 "2"^^ . _:b582918560 . _:b582919179 "2"^^ . _:b582919062 . _:b582918563 . _:b582919178 "2"^^ . . _:b582918562 . _:b582919189 "2"^^ . _:b582919213 . _:b582918439 . _:b582919188 "2"^^ . _:b582918573 . . _:b582918572 . _:b582919191 "2"^^ . _:b582919186 . _:b582918575 . _:b582919190 "2"^^ . _:b582918458 . _:b582918574 . _:b582919185 "2"^^ . _:b582918569 . _:b582919184 "2"^^ . _:b582918568 . _:b582919187 "2"^^ . _:b582918420 . _:b582918800 . _:b582918571 . _:b582919186 "2"^^ . . . _:b42012380 "Recently, the tumor suppressor NF2 (Merlin in D. melanogaster) was revealed to promote LATS1/2 kinase activation without activating the intrinsic kinase activity of MST1/2 (Yin et al. >>2013<<). Furthermore, Merlin is already an established transcriptional target of Yorkie, the fly homolog of YAP/TAZ (Hamaratoglu et al. 2006)." . _:b582919197 "2"^^ . _:b582918570 . _:b582918581 . _:b582919196 "2"^^ . _:b582919042 . _:b582919185 . _:b582918487 . _:b582918580 . _:b582919199 "2"^^ . _:b582918583 . _:b582919198 "2"^^ . . _:b582918582 . _:b582919193 "2"^^ . . _:b582918577 . _:b582919192 "2"^^ . _:b582918576 . _:b582919195 "2"^^ . _:b582918699 . . _:b582918747 . _:b582918579 . _:b582919194 "2"^^ . _:b582918578 . _:b582919205 "2"^^ . . . _:b582919166 . _:b582918589 . _:b582919204 "2"^^ . . _:b582918588 . _:b42012395 "by the mammalian Hippo pathway appears to differ significantly from that in D. melanogaster, where Yorkie induces expression of expanded, merlin, four-jointed, or kibra (Cho et al. 2006; Hamaratoglu et al. 2006; Genevet et al. >>2010<<), but not warts (the fly homolog of LATS1/2), in dividing epithelial cells of the wing or eye disc (Jukam et al. 2013)." . _:b582919207 "2"^^ . _:b582918819 . _:b582918591 . _:b582919206 "2"^^ . _:b582918923 . . _:b582919201 "2"^^ . _:b582919149 . . _:b582918590 . _:b582918585 . _:b582919200 "2"^^ . . _:b582918584 . _:b582919203 "2"^^ . _:b582919031 . _:b582919008 . _:b582918587 . _:b582919202 "2"^^ . _:b582919205 . . _:b582918586 . _:b582919213 "2"^^ . . _:b582918791 . _:b582918597 . _:b582919212 "2"^^ . _:b582918596 . _:b582919215 "2"^^ . _:b582918842 . _:b582918599 . _:b582919214 "2"^^ . . _:b582918743 . . _:b582918696 . _:b582918598 . _:b582919209 "2"^^ . . _:b582918926 . _:b582919236 . _:b582918593 . _:b582919208 "2"^^ . . _:b582919192 . _:b582918592 . _:b582919211 "2"^^ . _:b582919047 . . . . _:b582918416 . _:b582918680 . _:b582919210 "2"^^ . _:b582918595 . . _:b582918427 . _:b582919221 "2"^^ . _:b582918594 . . _:b582918605 . _:b582919220 "2"^^ . _:b582918414 "25"^^ . _:b582918604 . _:b582919223 "2"^^ . _:b582918415 "24"^^ . _:b582918692 . _:b582918705 . _:b582918413 "27"^^ . _:b582919222 "2"^^ . _:b582918607 . . _:b582918606 . _:b582919217 "2"^^ . _:b582918412 "30"^^ . _:b582918610 . _:b582919216 "2"^^ . _:b582918601 . _:b582919194 . _:b582918600 . _:b582919219 "2"^^ . . _:b582918899 . _:b582918416 "22"^^ . _:b582918603 . _:b582919218 "2"^^ . _:b582918642 . _:b582918419 "20"^^ . . _:b582918417 "22"^^ . . _:b582919229 "2"^^ . _:b582918602 . . _:b582919181 . _:b582918418 "21"^^ . _:b582918421 "18"^^ . _:b582918613 . _:b582919228 "2"^^ . _:b582918410 "40"^^ . _:b582918736 . _:b582919190 . . _:b582919231 "2"^^ . _:b582918905 . _:b582918612 . _:b582918420 "19"^^ . . . . _:b582919230 "2"^^ . . _:b582918615 . _:b582918422 "17"^^ . _:b582918411 "30"^^ . _:b582918614 . _:b582919225 "2"^^ . _:b582918601 . _:b582918423 "17"^^ . _:b582918609 . _:b582919224 "2"^^ . _:b582918431 "13"^^ . _:b582918424 "17"^^ . _:b582918608 . _:b582919227 "2"^^ . _:b582918425 "16"^^ . _:b582918430 "13"^^ . _:b582918491 . _:b582919226 "2"^^ . _:b582918611 . _:b582918426 "15"^^ . _:b582918424 . _:b582918427 "15"^^ . _:b582918610 . _:b582918428 "14"^^ . . _:b582919236 "2"^^ . _:b582918621 . _:b582918429 "14"^^ . . _:b582918620 . _:b582919004 . _:b582918623 . . . . _:b582918850 . _:b582919027 . . _:b582919233 "2"^^ . _:b582918622 . . _:b582919232 "2"^^ . _:b582918617 . _:b582918439 "12"^^ . _:b582918929 . _:b582918616 . _:b582919235 "2"^^ . _:b582918438 "13"^^ . . _:b42012367 . _:b582918559 . _:b582918735 . _:b582918619 . _:b582919234 "2"^^ . _:b582918437 "13"^^ . _:b582918618 . . _:b582918436 "13"^^ . _:b582918817 . _:b582918629 . . _:b582918435 "13"^^ . . _:b582918628 . _:b582918434 "13"^^ . _:b582918861 . _:b582918631 . _:b582918433 "13"^^ . _:b582918677 . _:b582918630 . . _:b582918432 "13"^^ . _:b582918527 . _:b582918625 . _:b582918447 "12"^^ . _:b582919225 . _:b582918624 . _:b582918446 "12"^^ . _:b582918627 . . _:b582918445 "12"^^ . _:b582918626 . _:b582918444 "12"^^ . . _:b582918980 . _:b582918637 . _:b582918443 "12"^^ . _:b582918615 . . _:b582918636 . _:b582918442 "12"^^ . _:b582918639 . _:b582918441 "12"^^ . . _:b582918760 . _:b582918638 . . . _:b582918440 "12"^^ . _:b582918455 "11"^^ . _:b582918633 . _:b582918454 "11"^^ . _:b582918632 . . _:b582918453 "11"^^ . _:b582918635 . _:b582918874 . _:b582919075 . _:b582918452 "11"^^ . _:b582918634 . . . _:b582918451 "11"^^ . . _:b582918645 . . _:b582918644 . _:b42012384 "To further confirm the role of LATS1/2 in feedback regulation, we generated LATS1/2-deficient mouse embryonic fibroblasts (MEFs) by infecting Lats1\u2013/\u2013Lats2F/F MEFs (Kim et al. >>2013<<) with adenoviruses encoding Cre recombinase." . . . _:b582918450 "12"^^ . _:b582919072 . _:b42012362 "To achieve liver-specific or intestine-specific gene deletion, YapF/F mice were bred to Albumin-Cre (Zhang et al. >>2010<<) or Villin-Cre (Taniguchi et al. 2015) Tg mice, respectively." . _:b582918647 . _:b582918449 "12"^^ . . . _:b582918646 . _:b582918448 "12"^^ . _:b582919079 . _:b582918889 . _:b582918463 "10"^^ . _:b582918641 . _:b582918869 . _:b582918462 "10"^^ . _:b582918640 . _:b582918938 . _:b582918461 "10"^^ . _:b582918825 . _:b582918565 . _:b582918643 . _:b582918460 "10"^^ . _:b582918642 . _:b582918459 "10"^^ . _:b582919158 . _:b582918653 . _:b582918661 . _:b582919073 . _:b582918562 . _:b582918458 "11"^^ . _:b582918652 . _:b582919134 . _:b582918457 "11"^^ . _:b582919161 . _:b582918639 . _:b582918655 . _:b582918456 "11"^^ . _:b582918654 . . _:b582918649 . _:b582918470 "9"^^ . _:b582918648 . _:b582918471 "9"^^ . . _:b582919165 . _:b582918651 . _:b582918468 "10"^^ . _:b582918650 . . _:b582918469 "9"^^ . _:b582918467 "10"^^ . _:b582918466 "10"^^ . _:b582918790 . _:b582919199 . _:b582918465 "10"^^ . _:b582918429 . _:b582919080 . _:b582918464 "10"^^ . . _:b582919139 . . _:b582918478 "9"^^ . . . . _:b582918479 "8"^^ . . _:b582919168 . . . . _:b582918476 "9"^^ . _:b582918810 . _:b582918477 "9"^^ . _:b582918517 . . _:b582918474 "9"^^ . _:b582919038 . . _:b42012385 "LPA acts through GPCR to induce YAP activation (Yu et al. >>2012<<). In control MCF10A cells, LPA treatment induced an acute YAP dephosphorylation, as determined by immunoblotting using the phospho-YAP antibody (Fig." . . _:b582918475 "9"^^ . . . _:b582918645 . _:b582918694 . _:b582918472 "9"^^ . _:b582918473 "9"^^ . . _:b582918576 . _:b582918486 "8"^^ . . . _:b582918487 "8"^^ . _:b582918484 "8"^^ . _:b582918799 . _:b582918485 "8"^^ . _:b582918482 "8"^^ . _:b582918483 "8"^^ . . . _:b582918847 . _:b582918480 "8"^^ . _:b582918678 . . _:b582919061 . . _:b582918481 "8"^^ . . _:b582918494 "8"^^ . _:b582918495 "8"^^ . . . _:b582918492 "8"^^ . _:b582918843 . _:b582918818 . _:b582918493 "8"^^ . . _:b582918490 "8"^^ . _:b582918491 "8"^^ . _:b582918488 "8"^^ . . _:b582919082 . _:b582918489 "8"^^ . _:b582918502 "7"^^ . . . _:b582918441 . _:b582918503 "7"^^ . _:b582918500 "7"^^ . _:b582918753 . _:b582919109 . _:b582918807 . . _:b582918663 . _:b582918501 "7"^^ . . . _:b582918498 "7"^^ . _:b582919227 . . _:b582918499 "7"^^ . _:b582918496 "7"^^ . . . _:b582918497 "7"^^ . . _:b582918510 "7"^^ . . _:b582918511 "7"^^ . _:b582919231 . _:b582918641 . _:b582918508 "7"^^ . _:b582918839 . . _:b582918509 "7"^^ . . _:b582918978 . _:b582918506 "7"^^ . "10.1101%2Fgad.262816.115" . . _:b582918507 "7"^^ . . _:b582918504 "7"^^ . . _:b582918505 "7"^^ . . _:b582918664 . _:b582918518 "6"^^ . . _:b582918519 "6"^^ . _:b582918739 . _:b582918516 "6"^^ . . _:b582918517 "6"^^ . _:b582918514 "6"^^ . _:b582918515 "6"^^ . . . _:b582918558 . _:b582918512 "7"^^ . _:b582918551 . _:b582918701 . _:b582918513 "6"^^ . . _:b582918526 "6"^^ . . _:b582918657 . _:b582918527 "6"^^ . . _:b582918524 "6"^^ . _:b582918525 "6"^^ . _:b582918522 "6"^^ . _:b582918523 "6"^^ . . . _:b582918520 "6"^^ . _:b582919087 . _:b582918521 "6"^^ . _:b582918788 . . _:b582918534 "6"^^ . _:b582918535 "6"^^ . . _:b582918532 "6"^^ . _:b582918877 . _:b582918533 "6"^^ . . _:b582918530 "6"^^ . _:b582918892 . _:b582918904 . _:b582918531 "6"^^ . _:b582918445 . _:b42012382 . _:b582919105 . _:b582918528 "6"^^ . _:b582918838 . . _:b582918529 "6"^^ . _:b582918797 . _:b582918542 "6"^^ . . . _:b582918543 "6"^^ . . _:b582918540 "6"^^ . _:b582919177 . . _:b582918541 "6"^^ . _:b582918695 . _:b582918538 "6"^^ . _:b582918539 "6"^^ . _:b582918536 "6"^^ . . . _:b582918575 . _:b582918537 "6"^^ . . . _:b582918549 "5"^^ . _:b582918548 "5"^^ . _:b582918551 "5"^^ . _:b582918927 . _:b42012364 . . _:b582918550 "5"^^ . _:b582918775 . _:b42012369 "of LATS1/2 individually, we immunoprecipitated endogenous LATS1 or LATS2 from MCF10A cells and examined the phosphorylation levels of their activation loop (AL) and HM, both of which correlate with LATS kinase activity (Hergovich >>2013<<). Immunoblot analysis revealed that activating phosphorylation of both LATS1 and LATS2 were markedly increased in cells overexpressing TAZ(4SA) (Fig." . _:b582918545 "5"^^ . _:b582918769 . _:b582918770 . . _:b582918544 "5"^^ . . _:b582918547 "5"^^ . _:b582919217 . . _:b582918546 "5"^^ . _:b582918557 "5"^^ . _:b582918556 "5"^^ . . _:b582918559 "5"^^ . . _:b582918442 . _:b582918558 "5"^^ . _:b582918941 . _:b582918805 . _:b582918553 "5"^^ . . _:b582918552 "5"^^ . . . _:b582918450 . _:b582918555 "5"^^ . . _:b582918671 . _:b582918554 "5"^^ . . . _:b582918565 "5"^^ . _:b582918564 "5"^^ . _:b582918567 "5"^^ . _:b582918436 . . . _:b582918566 "5"^^ . _:b582919146 . . _:b582918561 "5"^^ . _:b582918942 . . _:b582918484 . _:b582918560 "5"^^ . _:b582918802 . . . _:b582918563 "5"^^ . . . _:b582918562 "5"^^ . _:b582918856 . . . _:b582918573 "5"^^ . _:b582918572 "5"^^ . _:b582919068 . _:b582918575 "5"^^ . _:b582918574 "5"^^ . . _:b42012402 "Notably, the increase of AMOT may also contribute to the negative feedback because AMOT can directly bind to and inhibit YAP/TAZ as well as indirectly activate LATS (Zhao et al. 2011; Hirate et al. >>2013<<). One can predict with certainty that LATS must have many additional physiological substrates. Sequence analyses of the human proteome show that there are >1000 peptides with the LATS phosphorylation consensus motif (HXRXXS). Therefore," . _:b582918467 . _:b582918569 "5"^^ . _:b582918509 . _:b582918568 "5"^^ . . _:b582918508 . _:b582918571 "5"^^ . . _:b582918570 "5"^^ . _:b42012358 . _:b582918581 "5"^^ . _:b582918813 . _:b42012359 . _:b582918472 . _:b582918580 "5"^^ . . _:b582918583 "5"^^ . _:b582918599 . _:b582918582 "5"^^ . . _:b582918481 . _:b582918577 "5"^^ . _:b582918576 "5"^^ . _:b582919221 . _:b582918730 . _:b42012364 . _:b582918579 "5"^^ . _:b582918761 . _:b42012365 . _:b582918578 "5"^^ . _:b582919136 . _:b42012366 . _:b582918589 "5"^^ . _:b582918913 . _:b42012367 . _:b582918588 "5"^^ . _:b42012360 . _:b582918591 "5"^^ . _:b582918513 . _:b42012361 . _:b582918590 "5"^^ . . _:b582919032 . _:b42012407 "feedback regulators are found in human cancer: NF2 is mutated with high frequency in neurofibromatosis (Asthagiri et al. 2009), and loss of NF2 as well as LATS2 is also frequently observed in malignant mesothelioma (Murakami et al. >>2011<<), indicative of their functions as bona fide tumor suppressor genes." . _:b42012389 "While YAP and TAZ are known to promote cell migration (Chan et al. 2008; Haskins et al. 2014), recent studies revealed that the phosphorylation of AMOT by LATS1/2 inhibits AMOT binding to F-actin (Chan et al. 2013; Dai et al. >>2013<<; Hirate et al. 2013), reduces actin stress fiber formation, and thereby inhibits migration (Dai et al. 2013)." . _:b42012362 . _:b582918585 "5"^^ . _:b582918523 . _:b42012363 . . _:b582918584 "5"^^ . _:b42012372 . _:b42012389 . _:b582918587 "5"^^ . _:b42012373 . _:b582918586 "5"^^ . _:b582918648 . _:b42012374 . _:b42012391 . . _:b582918597 "5"^^ . _:b582918549 . _:b42012375 . _:b582918596 "5"^^ . _:b42012368 . _:b582918630 . _:b582918599 "5"^^ . . _:b42012369 . _:b582918598 "5"^^ . _:b42012370 . _:b582918593 "5"^^ . . _:b42012371 . _:b582918592 "5"^^ . . _:b42012380 . _:b582918595 "5"^^ . _:b582919191 . . _:b42012381 . _:b582918594 "5"^^ . _:b582918605 "4"^^ . _:b42012382 . _:b582918604 "4"^^ . _:b42012383 . _:b582918766 . _:b582918607 "4"^^ . _:b42012376 . . _:b582919129 . _:b582918606 "4"^^ . _:b42012377 . . _:b582918772 . _:b582918581 . _:b582918601 "4"^^ . _:b42012378 . . . _:b42012379 . _:b582918600 "5"^^ . _:b582918603 "4"^^ . _:b42012388 . . . _:b582918602 "4"^^ . _:b42012389 . _:b42012372 . . _:b582918613 "4"^^ . _:b42012390 . . . _:b582918612 "4"^^ . _:b42012391 . _:b582918545 . _:b582918937 . _:b582919202 . _:b582918615 "4"^^ . _:b42012384 . _:b582918614 "4"^^ . _:b42012385 . . . _:b582918609 "4"^^ . _:b42012386 . _:b582919057 . _:b582918608 "4"^^ . _:b42012387 . . . _:b582919091 . _:b582918611 "4"^^ . _:b42012396 . _:b582918732 . . _:b582918610 "4"^^ . _:b42012397 . . . . _:b582918621 "4"^^ . _:b42012398 . _:b582918620 "4"^^ . _:b42012399 . _:b582918623 "4"^^ . _:b42012392 . _:b582918622 "4"^^ . _:b42012393 . _:b582919137 . _:b582918617 "4"^^ . _:b42012394 . . _:b582918616 "4"^^ . _:b42012395 . _:b582918619 "4"^^ . _:b42012404 . _:b582918713 . _:b582918963 . . _:b582918483 . _:b42012405 . _:b582918618 "4"^^ . _:b582918910 . _:b582919017 . . . _:b582918767 . _:b42012406 . _:b582918629 "4"^^ . . _:b582918628 "4"^^ . _:b42012407 . _:b582918586 . _:b582918631 "4"^^ . _:b42012400 . . _:b582918630 "4"^^ . _:b42012401 . . . _:b582918625 "4"^^ . _:b42012402 . . _:b582919043 . . _:b582918750 . _:b42012403 . _:b582918624 "4"^^ . _:b582919119 . _:b582918627 "4"^^ . _:b582918626 "4"^^ . . _:b582919114 . _:b42012366 "TEAD is the major YAP-associated transcription factor, and its binding to YAP requires the Ser94 residue in YAP (Zhao et al. >>2008<<). Mutating Ser94 abolished the ability of YAP(5SA) to suppress TAZ (Fig. 1D). Consistently, shRNA-mediated depletion of endogenous TEAD1/3/4 (Supplemental Fig. S1B) attenuated the effect of YAP(5SA) on TAZ (Fig. 1E), suggesting that the" . _:b582918637 "4"^^ . _:b582919224 . _:b582918636 "4"^^ . _:b582918639 "4"^^ . _:b582918660 . _:b582918638 "4"^^ . _:b582918617 . _:b582918633 "4"^^ . _:b582918632 "4"^^ . _:b582918561 . _:b582919033 . . _:b582918635 "4"^^ . . . _:b582918634 "4"^^ . _:b582918645 "4"^^ . _:b582918422 . _:b582918477 . _:b582918644 "4"^^ . . _:b582918647 "4"^^ . _:b582918646 "4"^^ . . _:b582918616 . _:b582918641 "4"^^ . _:b582918640 "4"^^ . _:b582918550 . _:b582918928 . _:b582918455 . _:b582918643 "4"^^ . _:b582919036 . . _:b582918642 "4"^^ . _:b582918900 . _:b582918653 "4"^^ . _:b582918652 "4"^^ . _:b582919180 . _:b582918655 "4"^^ . _:b582918704 . _:b582918654 "4"^^ . _:b582918649 "4"^^ . _:b582918708 . _:b582919107 . . _:b582919095 . _:b582918648 "4"^^ . _:b582918729 . _:b582918885 . _:b582918651 "4"^^ . _:b582918921 . _:b582918589 . _:b582919001 . _:b582918650 "4"^^ . . _:b582918566 . . _:b582919187 . . . _:b42012396 "expression of expanded, merlin, four-jointed, or kibra (Cho et al. 2006; Hamaratoglu et al. 2006; Genevet et al. 2010), but not warts (the fly homolog of LATS1/2), in dividing epithelial cells of the wing or eye disc (Jukam et al. >>2013<<). On the contrary, Yorkie indirectly suppresses Warts transcription to establish positive feedback regulation in combination with tissue-specific transcription factors in the context of Drosophila eye development (Jukam et al. 2013)." . _:b582918768 . . _:b582918432 . _:b582918603 . _:b582918863 . _:b582919147 . _:b582919020 . _:b582918984 . . _:b582918703 . _:b582919179 . _:b582918644 . . . _:b582919219 . _:b582918568 . . . . . _:b582919169 . . _:b582918437 . . _:b582919117 . _:b582919128 . _:b582919102 . . _:b582918893 . _:b582918965 . _:b582918585 . _:b582918947 . _:b42012380 . _:b582918960 . . _:b582918757 . _:b582919173 . _:b582919172 . _:b582919175 . _:b582919174 . . _:b582919067 . _:b582919169 . . _:b582919168 . _:b582918895 . _:b582919171 . . _:b582919170 . . _:b582919181 . _:b582918588 . . _:b582919180 . _:b582919183 . _:b582919182 . _:b582919177 . . . _:b582919176 . _:b582918779 . _:b582919179 . _:b582919132 . _:b582919106 . _:b582919178 . _:b582919189 . . . _:b582918553 . _:b582919188 . . . . _:b582919071 . _:b582918679 . _:b582919191 . . "PMC0" . _:b582918811 . . _:b582919190 . _:b582918532 . _:b582919185 . _:b582918748 . . _:b582919184 . . _:b42012398 . _:b582919187 . . _:b582919186 . _:b42012388 "While YAP and TAZ are known to promote cell migration (Chan et al. 2008; Haskins et al. 2014), recent studies revealed that the phosphorylation of AMOT by LATS1/2 inhibits AMOT binding to F-actin (Chan et al. >>2013<<; Dai et al. 2013; Hirate et al. 2013), reduces actin stress fiber formation, and thereby inhibits migration (Dai et al. 2013)." . _:b582918725 . _:b582919197 . . _:b582919196 . . _:b582919199 . _:b582918528 . . _:b582918801 . _:b582919198 . _:b582919193 . _:b582919192 . _:b582918465 . _:b582918534 . . _:b582919195 . _:b42012403 "Given the observed frequent YAP/TAZ activation in human cancers, the general lack of cancer-relevant somatic or germline mutations in core members of the Hippo pathway is a major conundrum to the field (Harvey et al. >>2013<<; Moroishi et al. 2015; Plouffe et al. 2015). One possibility is that negative feedback control of YAP/TAZ activity suppresses tumorigenesis following small perturbation of the pathway." . _:b582919194 . _:b582919205 . _:b582918466 . _:b582918718 . _:b582919204 . _:b582919124 . _:b582919207 . . _:b582919206 . . . _:b582919201 . _:b582919200 . _:b582919056 . _:b582919203 . . . _:b582919202 . _:b582918784 . _:b582919213 . _:b42012404 "the observed frequent YAP/TAZ activation in human cancers, the general lack of cancer-relevant somatic or germline mutations in core members of the Hippo pathway is a major conundrum to the field (Harvey et al. 2013; Moroishi et al. >>2015<<; Plouffe et al. 2015). One possibility is that negative feedback control of YAP/TAZ activity suppresses tumorigenesis following small perturbation of the pathway." . . _:b582918627 . _:b582919212 . . . . _:b582919215 . _:b582919101 . _:b582919214 . . _:b582919209 . _:b582919208 . _:b582918831 . _:b582918964 . _:b582919211 . . _:b582918596 . _:b582919210 . _:b582919221 . _:b582919220 . . _:b582919223 . . _:b582919222 . _:b582919217 . _:b582919216 . _:b582918646 . _:b582918530 . _:b582919219 . . _:b582919218 . _:b582918783 . _:b582919229 . _:b42012370 "Consistent with the observed LATS1 and LATS2 activation, phosphorylation of angiomotin (AMOT) at Ser175, a known direct substrate site for LATS1/2 kinases (Adler et al. >>2013<<; Chan et al. 2013; Dai et al. 2013; Hirate et al. 2013), was increased in TAZ(4SA)-overexpressing MCF10A cells (Fig." . _:b582919228 . _:b582919098 . . _:b582919231 . . _:b582919230 . _:b582918835 . _:b582919225 . _:b582918414 . _:b582918676 . _:b582919224 . . _:b582919227 . . . _:b582919226 . _:b42012366 . _:b582919236 . . _:b582919215 . _:b582919063 . _:b582918480 . _:b582919233 . . _:b582918931 . _:b582919189 . _:b582919232 . . . _:b582919235 . _:b582918464 . _:b582919234 . _:b582918622 . _:b582918415 . . . . _:b582918459 . _:b582919235 . _:b582918670 . . . _:b582918884 . . _:b582918635 . _:b582918564 . _:b582918981 . . _:b582919007 . . _:b582919003 . _:b582918738 . _:b42012376 . . _:b582918507 . . _:b582918675 . _:b582918902 . _:b582918845 . _:b582918529 . _:b582919044 . . _:b582918658 . _:b582918500 . _:b582919093 . . _:b42012400 "uncoupling of the GPCR from the G proteins (Pierce et al. 2002) or feedback phosphorylation of upstream components of a signaling cascade (e.g., inhibitory phosphorylation of RAF by ERK in the MAPK signaling) (Dougherty et al. >>2005<<). Thus, the LATS-mediated feedback architecture provides enough time for YAP/TAZ to mediate their biological functions as transcriptional coactivators by taking advantage of the time delay of transcription and translation." . _:b582919212 . _:b582919213 . _:b582919214 . _:b582919215 . _:b582919208 . _:b582919209 . _:b582919210 . _:b582919211 . _:b582919204 . _:b582919205 . _:b42012365 _:b42012368 . _:b582919206 . _:b42012365 _:b42012369 . . _:b42012365 _:b42012370 . _:b582919207 . _:b582919200 . _:b42012365 _:b42012371 . _:b582919201 . . _:b42012365 _:b42012372 . _:b582919202 . _:b42012365 _:b42012373 . _:b582919203 . _:b42012365 _:b42012374 . _:b582919228 . _:b42012365 _:b42012375 . _:b582919229 . _:b42012365 _:b42012376 . _:b582919230 . _:b42012365 _:b42012377 . _:b582919231 . _:b582918911 . _:b42012365 _:b42012378 . _:b582919224 . _:b42012365 _:b42012379 . _:b582919225 . _:b582919150 . _:b42012365 _:b42012380 . _:b582919226 . _:b42012365 _:b42012381 . _:b582919227 . _:b42012365 _:b42012382 . _:b582919220 . _:b42012365 _:b42012383 . . _:b582919221 . _:b582919222 . _:b582919223 . _:b582919216 . . _:b582919217 . _:b582919218 . _:b582919219 . _:b582919180 . _:b582919181 . _:b582919182 . . _:b582919183 . _:b582919176 . _:b582918709 . _:b582919177 . _:b582919178 . _:b582918731 . _:b42012365 _:b42012366 . _:b582919179 . _:b582919172 . _:b42012365 _:b42012367 . _:b582919173 . _:b582919174 . _:b582919175 . _:b582919168 . _:b582919169 . _:b582919170 . _:b582919171 . _:b582919196 . _:b582919197 . . _:b582919198 . _:b582919199 . _:b582919192 . _:b582918688 . . _:b582919193 . _:b582919194 . . _:b582919195 . _:b582919188 . _:b582919189 . _:b42012365 _:b42012384 . _:b42012358 _:b42012364 . _:b582919190 . _:b42012365 _:b42012385 . _:b582919191 . _:b42012365 _:b42012386 . _:b582919184 . _:b42012365 _:b42012387 . _:b582919185 . _:b42012358 _:b42012360 . _:b42012365 _:b42012388 . _:b582919186 . _:b42012365 _:b42012389 . _:b42012358 _:b42012361 . _:b582919187 . _:b42012365 _:b42012390 . _:b42012358 _:b42012362 . _:b42012365 _:b42012391 . _:b42012358 _:b42012363 . . _:b42012358 _:b42012359 . _:b582918594 . . . . . _:b582918919 . _:b582919048 . _:b582919153 . _:b42012371 "Consistent with the observed LATS1 and LATS2 activation, phosphorylation of angiomotin (AMOT) at Ser175, a known direct substrate site for LATS1/2 kinases (Adler et al. 2013; Chan et al. >>2013<<; Dai et al. 2013; Hirate et al. 2013), was increased in TAZ(4SA)-overexpressing MCF10A cells (Fig." . . _:b582918979 . _:b582919236 . _:b582919232 . _:b582919233 . _:b582919143 . _:b582919234 . _:b582919235 . _:b582919064 . . . . _:b582919152 . . _:b42012374 "These results are consistent with the previous observation that LATS-dependent phosphorylation stabilizes AMOT (Adler et al. >>2013<<). We also determined endogenous YAP localization in TAZ(4SA)-expressing cells. As expected, TAZ(4SA) overexpression caused a dramatic alteration of YAP subcellular distribution in MCF10A cells (Fig. 2F). There was much more cytoplasmic" . . . _:b582918482 . . _:b582918505 . _:b582919151 . . _:b582918563 . . _:b582918683 . . _:b582918901 . _:b582918796 . . _:b582918502 . _:b582918959 . _:b582918419 . _:b582919173 . _:b582918421 . _:b582918554 . . _:b42012392 _:b42012396 . _:b42012392 _:b42012397 . _:b42012392 _:b42012398 . _:b42012392 _:b42012399 . _:b582918468 . _:b42012392 _:b42012393 . _:b42012392 _:b42012394 . _:b42012392 _:b42012395 . _:b582918933 . _:b42012392 _:b42012404 . _:b42012392 _:b42012405 . _:b42012392 _:b42012406 .