. . . . _:b584504066 . _:b584504006 "2"^^ . _:b584504001 "2"^^ . _:b584503847 . _:b584504000 "2"^^ . _:b42685562 . _:b584504041 . . _:b584504003 "2"^^ . _:b584503781 . . _:b584503968 . . . _:b584504002 "2"^^ . _:b584504008 . _:b584503790 . _:b584503784 . _:b584503902 . _:b584504013 "2"^^ . _:b584503741 . . _:b584504012 "2"^^ . . . _:b584504015 "2"^^ . _:b42685571 . _:b42685537 . _:b584504014 "2"^^ . _:b42685524 . _:b584504009 "2"^^ . _:b584504093 . _:b584503956 . _:b584504155 . _:b584504042 . _:b584504008 "2"^^ . _:b584503971 . . _:b584504011 "2"^^ . _:b584503942 . _:b584504010 "2"^^ . _:b42685549 . _:b584503882 . . _:b584504021 "2"^^ . _:b584503793 . _:b584504020 "2"^^ . _:b584504023 "2"^^ . _:b584504084 . _:b584504022 "2"^^ . . _:b584504017 "2"^^ . _:b42685563 . _:b584503720 . _:b584504016 "2"^^ . . . _:b584504019 "2"^^ . . _:b42685529 . _:b584504018 "2"^^ . . . _:b584504029 "2"^^ . _:b584503761 . _:b584504028 "2"^^ . . _:b42685553 . _:b584504031 "2"^^ . _:b584504030 "2"^^ . _:b584504025 "2"^^ . _:b584504024 "2"^^ . . _:b584504027 "2"^^ . . _:b42685556 "These data suggest that RFS-1/RIP-1 physically associates with RAD-51-ssDNA filaments, similar to the budding yeast Rad55-Rad57 and Shu complexes (Liu et\u00A0al., 2011a; Sasanuma et\u00A0al., >>2013<<)." . _:b584503910 . _:b42685572 . _:b42685552 "rip-1 mutants also phenocopy the meiotic HR defects of rfs-1 strains (Ward et\u00A0al., >>2010<<), exhibiting elevated frequencies of males (Figure\u00A0S1G) and synthetic lethality with helq-1, which is associated with persistent meiotic RAD-51 foci (Figures S1H\u2013S1J)." . . _:b584504026 "2"^^ . . . _:b584504037 "2"^^ . . _:b584504102 . _:b584504036 "2"^^ . _:b42685560 . _:b42685563 "Since Rad51 focus formation after damage is also defective in Rad51 paralog mutants from other eukaryotic organisms (Gasior et\u00A0al., >>1998<<), the filament remodeling function of RFS-1/RIP-1 to a stable conformation is likely to be conserved." . _:b584504039 "2"^^ . _:b584504038 "2"^^ . _:b42685553 . . . _:b584504033 "2"^^ . . . _:b584504032 "2"^^ . _:b584503966 . _:b584504035 "2"^^ . . _:b584503964 . _:b584503791 . _:b584503712 . _:b584504034 "2"^^ . _:b584503755 . _:b584504045 "2"^^ . . _:b584503794 . _:b584504044 "2"^^ . _:b584504047 "2"^^ . _:b584503896 . _:b584504157 . _:b584503985 . _:b584504046 "2"^^ . . _:b584504041 "2"^^ . _:b584503737 . _:b584504040 "2"^^ . _:b584504030 . _:b584504031 . _:b584504043 "2"^^ . _:b584504006 . _:b42685547 . _:b584503758 . _:b42685569 . _:b584504042 "2"^^ . . . . . _:b584504053 "2"^^ . . _:b584503759 . _:b42685536 "family reflects their functional importance in conferring resistance to DNA damage and the integrity of the Rad51 paralog complexes in mammalian cells (French et\u00A0al., 2003; Gruver et\u00A0al., 2005; Wiese et\u00A0al., 2006; Yamada et\u00A0al., >>2004<<)." . _:b584504052 "2"^^ . . . _:b584504055 "2"^^ . _:b584503999 . . _:b42685551 . _:b584504054 "2"^^ . _:b584504049 "2"^^ . _:b584503778 . . _:b584504048 "2"^^ . . _:b584504027 . _:b584504051 "2"^^ . _:b584504024 . _:b42685558 "All these observations are in striking contrast to Srs2, which disassembles Rad51-ssDNA filaments, inhibits Rad51 recombinase activity, and antagonizes Rad51 focus formation in yeast (Burgess et\u00A0al., 2009; Krejci et\u00A0al., >>2003<<). We therefore favor the second model, in which the reduction in fluorescence is attributed to a change in the biophysical properties (hereafter referred to as \u201Cremodeling\u201D) of the RAD-51-ssDNA filaments." . . _:b584504050 "2"^^ . _:b584504061 "2"^^ . . . . _:b584504060 "2"^^ . _:b584504106 . _:b42685537 . _:b584504063 "2"^^ . _:b584503945 . _:b42685530 . _:b584504090 . . _:b42685550 "Since Walker B motifs mediate protein-protein interactions in human Rad51 paralog complexes (Wiese et\u00A0al., >>2006<<), we tested if this dependency is conserved for the RFS-1/RIP-1 protein-protein interaction." . _:b42685547 "We previously showed that rfs-1 mutant strains are sensitive to DNA damage, defective for RAD-51 focus formation at stalled replication forks, and exhibit meiotic defects when combined with helq-1 mutations (Ward et\u00A0al., 2007, >>2010<<). However, attempts to purify RFS-1 yielded largely insoluble protein intractable to biochemical analysis (Figure\u00A0S1A). Since Rad51 paralogs in other organisms function as complexes, we reasoned that RFS-1 may require a binding partner" . _:b584504062 "2"^^ . . . _:b584504057 "2"^^ . _:b584503967 . . _:b584504112 . _:b584504056 "2"^^ . . . _:b584503760 . _:b584504059 "2"^^ . _:b584503716 . _:b584503926 . _:b42685533 . _:b584504058 "2"^^ . _:b584503939 . _:b584503799 . _:b42685538 . . _:b584504069 "2"^^ . . _:b584504068 "2"^^ . _:b42685535 . _:b584504069 . . _:b584504071 "2"^^ . . _:b42685567 . _:b42685526 . . _:b584504070 "2"^^ . _:b584503735 . _:b584504065 "2"^^ . . _:b584504064 "2"^^ . _:b42685564 "This result is also consistent with the suppression of the DNA damage sensitivity and HR defects of yeast rad55\u0394 and rad57\u0394 mutants by a mutant form of Rad51 (I345T) that binds DNA more stably (Fortin and Symington, >>2002<<)." . _:b42685525 . . _:b584504067 "2"^^ . . _:b584504001 . . . _:b584504066 "2"^^ . . _:b584504150 . _:b584504077 "2"^^ . _:b42685555 "The nematode RAD-51 is an extremely weak recombinase in the absence of mediators even when tested at a wide range of protein-to-DNA ratios (Petalcorin et\u00A0al., >>2006<<). Nevertheless, addition of sub-stoichiometric concentrations of RFS-1/RIP-1 relative to RAD-51 caused a dramatic stimulation of both D loop formation (Figure\u00A02C) and strand exchange activities (Figure\u00A0S2D), which was dependent on the" . _:b584503738 . _:b584504076 "2"^^ . _:b584504079 "2"^^ . _:b42685538 . . _:b584504078 "2"^^ . _:b42685572 . . _:b584503804 . _:b584503807 . _:b584504073 "2"^^ . _:b584503885 . _:b584504025 . _:b584504072 "2"^^ . . _:b584503980 . . _:b584504075 "2"^^ . _:b584503874 . . _:b584504026 . _:b584504074 "2"^^ . . _:b584504085 "2"^^ . _:b584504084 "2"^^ . _:b584504087 "2"^^ . . _:b584503923 . _:b42685537 "at an early stage in the HR reaction (Chun\u00A0et\u00A0al., 2013; French et\u00A0al., 2002; Gasior et\u00A0al., 1998; Godthelp et\u00A0al., 2002; Hays et\u00A0al., 1995; Johnson et\u00A0al., 1999; Pierce et\u00A0al., 1999; Rattray and Symington, 1995; Takata et\u00A0al., 2000, >>2001<<). Like BRCA2 and PALB2, which are mutated in Fanconi anemia and breast and ovarian cancer (Howlett et\u00A0al., 2002; Lancaster et\u00A0al., 1996; Rahman et\u00A0al., 2007; Reid et\u00A0al., 2007; Wooster et\u00A0al., 1995; Xia et\u00A0al., 2007), biallelic germline" . . _:b584504086 "2"^^ . _:b42685530 "paralog proteins share sequence similarity to Rad51 and possess homology extending across the motor ATPase fold, including the highly conserved Walker A and Walker B boxes, but not the N-terminal helix-hairpin-helix motif (Lin et\u00A0al., >>2006<<). Five Rad51 paralogs have been identified in mammalian and avian species, which interact with one another to form two constitutive complexes, RAD51B-RAD51C-RAD51D-XRCC2 (BCDX2) and RAD51C-XRCC3 (CX3) (Masson et\u00A0al., 2001; Yonetani et\u00A0al." . _:b584504081 "2"^^ . . _:b584504109 . . _:b584504080 "2"^^ . _:b584504059 . _:b584504083 "2"^^ . . _:b584504082 "2"^^ . _:b584503813 . _:b584503921 . _:b584504093 "2"^^ . _:b584503812 . _:b584504092 "2"^^ . _:b584503815 . _:b584504147 . . _:b584504095 "2"^^ . . _:b584504159 . _:b584503814 . . _:b584504094 "2"^^ . _:b584503809 . _:b584503950 . _:b584504046 . . _:b584504089 "2"^^ . _:b584503808 . . _:b584504088 "2"^^ . _:b584503811 . _:b584504091 "2"^^ . . _:b584503810 . _:b584504090 "2"^^ . _:b42685543 "Rad55-Rad57 and the Shu complex form co-complexes with Rad51-ssDNA filaments (Liu et\u00A0al., 2011a; Sasanuma et\u00A0al., >>2013<<), which in the case of Rad55-Rad57 renders the pre-synaptic complex resistant to disruption by the anti-recombinase Srs2 (Liu et\u00A0al., 2011a)." . _:b584503821 . _:b42685554 . . _:b584504101 "2"^^ . _:b42685551 "DNA-damaging agents that stall replication forks (Figure\u00A01D), are sensitive to DNA damage (Figures S1D and S1E), and display elevated germline apoptosis after treatment with interstrand crosslinking agents (Figure\u00A0S1F) (Ward et\u00A0al., >>2007<<). rip-1 mutants also phenocopy the meiotic HR defects of rfs-1 strains (Ward et\u00A0al., 2010), exhibiting elevated frequencies of males (Figure\u00A0S1G) and synthetic lethality with helq-1, which is associated with persistent meiotic RAD-51 foci" . _:b584503820 . _:b584503765 . . . _:b584504100 "2"^^ . _:b584503823 . . _:b42685557 . _:b584503842 . _:b584504103 "2"^^ . . _:b584503822 . _:b584504038 . _:b584504102 "2"^^ . _:b584503817 . _:b584503978 . _:b584504097 "2"^^ . _:b584504033 . _:b584503816 . _:b584504096 "2"^^ . . _:b584503819 . _:b42685540 "anemia (Vaz et\u00A0al., 2010), whereas monoallelic inheritance of mutations in RAD51C and RAD51D, and RAD51B, predispose individuals to ovarian and breast cancer, respectively (Golmard et\u00A0al., 2013; Loveday et\u00A0al., 2011; Meindl et\u00A0al., >>2010<<), demonstrating an important tumor suppressor function for HR mediators." . _:b584503722 . _:b584504099 "2"^^ . _:b584503818 . . _:b584504098 "2"^^ . _:b584504123 . _:b584504060 . _:b584503829 . _:b584503873 . . _:b584504045 . _:b584504109 "2"^^ . _:b584503828 . _:b584504108 "2"^^ . _:b584503831 . _:b584504111 "2"^^ . _:b584503830 . _:b584504145 . _:b584504110 "2"^^ . . _:b584503825 . _:b42685537 . _:b584503988 . _:b584504105 "2"^^ . _:b584503824 . _:b584504104 "2"^^ . _:b584503827 . _:b584504107 "2"^^ . _:b584503826 . _:b584504106 "2"^^ . _:b42685537 . _:b584503837 . _:b584503937 . _:b584504117 "2"^^ . _:b584503836 . _:b584504116 "2"^^ . . _:b584504152 . _:b584503839 . . _:b584504119 "2"^^ . . _:b584503838 . _:b584504118 "2"^^ . . _:b584504010 . _:b584503797 . . _:b584503833 . . _:b584504113 "2"^^ . _:b584503832 . _:b584503990 . _:b584504112 "2"^^ . _:b584503835 . _:b42685528 "For example, BRCA2 facilitates Rad51 nuclear localization (Jeyasekharan et\u00A0al., 2013; Martin et\u00A0al., 2005), RPA displacement from ssDNA, and Rad51 filament nucleation (Jensen et\u00A0al., 2010; Liu et\u00A0al., 2010; Thorslund et\u00A0al., >>2010<<), whereas Rad54 unloads Rad51 from dsDNA to promote repair DNA synthesis (Solinger et\u00A0al., 2002)." . . . _:b584504115 "2"^^ . _:b584503834 . . _:b584504153 . _:b584504114 "2"^^ . _:b584503845 . _:b584504125 "2"^^ . _:b584503844 . . _:b584504124 "2"^^ . . _:b584503847 . _:b584503893 . _:b584503718 "9"^^ . _:b584504127 "2"^^ . _:b584503913 . . _:b584503846 . _:b42685524 "introduction" . _:b584503719 "8"^^ . . . _:b584504126 "2"^^ . . _:b584503841 . _:b584503716 "9"^^ . _:b584504121 "2"^^ . _:b584503840 . . _:b584503717 "9"^^ . _:b584504120 "2"^^ . _:b584503843 . _:b584504123 "2"^^ . _:b584503714 "10"^^ . _:b42685535 "to Rad51 but contain Rad51-like folds as determined by crystal structures (Sasanuma et\u00A0al., 2013; Tao et\u00A0al., 2012) or show very limited homology centered on the Walker B motif, such as budding/fission yeast Psy3/Rdl1 (Mart\u00EDn et\u00A0al., >>2006<<). The high degree of conservation around these short Walker motifs among the Rad51 paralog family reflects their functional importance in conferring resistance to DNA damage and the integrity of the Rad51 paralog complexes in mammalian" . _:b584503842 . _:b584503715 "9"^^ . _:b584504122 "2"^^ . _:b584503713 "11"^^ . _:b584503853 . _:b584504154 . _:b584504133 "2"^^ . . _:b584503906 . _:b584503852 . _:b584503712 "12"^^ . _:b584504132 "2"^^ . . _:b584503726 "7"^^ . _:b584503855 . _:b584503903 . _:b584504047 . _:b584504135 "2"^^ . _:b584503727 "6"^^ . _:b42685534 "fission yeast Shu complexes (Mart\u00EDn et\u00A0al., 2006; Shor et\u00A0al., 2005) contain proteins lacking detectable sequence homology to Rad51 but contain Rad51-like folds as determined by crystal structures (Sasanuma et\u00A0al., 2013; Tao et\u00A0al., >>2012<<) or show very limited homology centered on the Walker B motif, such as budding/fission yeast Psy3/Rdl1 (Mart\u00EDn et\u00A0al., 2006)." . _:b584503946 . _:b584503854 . _:b584504036 . _:b584504134 "2"^^ . _:b584503724 "7"^^ . _:b584503959 . _:b584503849 . . _:b584504129 "2"^^ . _:b584503725 "7"^^ . _:b584503848 . _:b584504128 "2"^^ . _:b584503862 . _:b584504005 . _:b584503722 "7"^^ . _:b584503851 . _:b584503826 . _:b584504131 "2"^^ . _:b584503723 "7"^^ . _:b584503850 . . _:b584504130 "2"^^ . _:b584503861 . _:b584503720 "8"^^ . _:b584504141 "2"^^ . _:b584504074 . _:b584503860 . _:b584503721 "8"^^ . _:b584504140 "2"^^ . . _:b584503734 "6"^^ . _:b584503863 . _:b584504143 "2"^^ . _:b584503735 "6"^^ . _:b584503862 . _:b584504142 "2"^^ . _:b584503732 "6"^^ . _:b584503857 . . _:b584504137 "2"^^ . _:b584503733 "6"^^ . . _:b584503856 . _:b584504136 "2"^^ . _:b584503730 "6"^^ . . _:b584503859 . . _:b42685537 . _:b584504139 "2"^^ . _:b584503731 "6"^^ . _:b584503858 . _:b584504138 "2"^^ . _:b584503728 "6"^^ . _:b584503869 . _:b584504149 "2"^^ . _:b42685537 . _:b584503729 "6"^^ . _:b42685524 . _:b584503868 . _:b584504148 "2"^^ . _:b42685525 . _:b584503871 . _:b42685557 "By monitoring changes in Cy3 fluorescence over time, we could quantitatively assess changes in the biophysical properties of the fluorophore as a proxy for changes in the nature of the protein-DNA association (Antony et\u00A0al., >>2009<<). Although fluorescence was stable in the absence of protein, rapid mixing of RAD-51 with the labeled DNA in the presence of ATP resulted in an increase in fluorescence as a function of time (Figure\u00A04A). Both the size and the rate of" . _:b584503741 "5"^^ . _:b42685526 . _:b584504151 "2"^^ . _:b584503713 . _:b584504003 . . _:b584503870 . _:b584503740 "5"^^ . _:b42685527 . _:b584504150 "2"^^ . . _:b584503865 . . _:b584503743 "5"^^ . _:b584504145 "2"^^ . . _:b584503864 . . . _:b584503742 "5"^^ . _:b584504144 "2"^^ . _:b584503738 "6"^^ . _:b584503867 . _:b584503880 . _:b584504147 "2"^^ . _:b584503739 "6"^^ . _:b584503866 . _:b584504146 "2"^^ . _:b584503736 "6"^^ . _:b584503979 . . _:b584503877 . _:b584504157 "2"^^ . _:b42685532 . _:b584503737 "6"^^ . . _:b584503876 . _:b584504156 "2"^^ . _:b42685533 . . _:b584504098 . _:b584504096 . _:b584503879 . _:b584503922 . _:b584503993 . _:b584503749 "5"^^ . _:b42685534 . _:b584504159 "2"^^ . _:b584503878 . _:b584503933 . . _:b42685535 . _:b584504158 "2"^^ . _:b584503748 "5"^^ . _:b584503873 . _:b584503751 "5"^^ . _:b42685528 . _:b584504153 "2"^^ . _:b584503872 . _:b584503750 "5"^^ . _:b42685529 . _:b584504152 "2"^^ . . _:b584503814 . _:b584503875 . . _:b584504104 . _:b584503745 "5"^^ . _:b42685530 . _:b584504155 "2"^^ . . _:b584503874 . _:b584503744 "5"^^ . _:b42685531 . _:b584504154 "2"^^ . . _:b584503885 . _:b584503747 "5"^^ . _:b42685540 . . _:b584503884 . . _:b584503834 . _:b584503746 "5"^^ . _:b42685541 . _:b584503887 . . _:b42685542 . _:b584503757 "5"^^ . _:b584504116 . _:b584503886 . _:b584503756 "5"^^ . _:b42685543 . _:b584503881 . _:b584503759 "5"^^ . _:b42685536 . . _:b584503880 . _:b584504081 . _:b584503758 "5"^^ . _:b42685537 . _:b42685528 . _:b584504160 "2"^^ . _:b42685528 . . _:b584503883 . . _:b584503753 "5"^^ . _:b42685538 . _:b584503912 . . _:b42685540 . _:b584503730 . . _:b584504075 . _:b584503882 . . _:b584503752 "5"^^ . _:b42685539 . . _:b584503893 . . _:b42685548 . . _:b584503755 "5"^^ . _:b584504070 . _:b584503892 . _:b584504018 . _:b584503853 . _:b584503754 "5"^^ . _:b42685549 . _:b584503843 . . _:b584503895 . _:b584503765 "4"^^ . _:b42685550 . . _:b584503894 . _:b584503803 . . _:b42685551 . _:b584503764 "5"^^ . . _:b42685572 "from that proposed for other positive regulators of HR, which are epistatic to Rad51 paralogs, including BRCA2 and Rad54 (Chun et\u00A0al., 2013; Jensen et\u00A0al., 2010, 2013; Liu et\u00A0al., 2010; Solinger et\u00A0al., 2002; Thorslund et\u00A0al., >>2010<<). We therefore propose a model (Figure\u00A07I) for HR, in which Rad51-ssDNA filaments are first nucleated by BRCA2, displacing RPA from ssDNA." . _:b584503889 . . _:b584503767 "4"^^ . _:b42685544 . _:b584503888 . _:b584503766 "4"^^ . . . _:b42685545 . . _:b584503891 . _:b584503761 "5"^^ . _:b42685546 . _:b584503863 . . . _:b584503890 . _:b42685531 . _:b584503760 "5"^^ . _:b42685547 . . . . _:b584503901 . _:b584503763 "5"^^ . _:b42685556 . _:b42685534 . . . _:b584503900 . _:b584503895 . _:b584503762 "5"^^ . _:b42685557 . . _:b584504097 . . _:b584503903 . _:b584503773 "4"^^ . _:b42685558 . . _:b584503902 . _:b584503772 "4"^^ . . _:b42685559 . . . . _:b584503897 . . _:b584503775 "4"^^ . _:b584503787 . _:b42685552 . _:b584503896 . _:b584503774 "4"^^ . _:b42685553 . . _:b584503899 . . _:b584503769 "4"^^ . _:b42685537 . _:b42685554 . . . _:b584503898 . _:b584503768 "4"^^ . . _:b42685555 . _:b584503789 . . _:b584503909 . _:b584503771 "4"^^ . _:b584504124 . _:b42685564 . . _:b584503908 . _:b584503770 "4"^^ . _:b584503861 . _:b42685565 . _:b584503911 . _:b584503781 "4"^^ . _:b42685566 . _:b584503910 . . _:b42685528 . _:b584503780 "4"^^ . _:b42685567 . _:b584503905 . _:b584503783 "4"^^ . _:b42685560 . _:b584503907 . _:b584503997 . . _:b584503904 . _:b584503782 "4"^^ . _:b584503767 . _:b42685568 . _:b42685561 . _:b584503907 . . _:b584503777 "4"^^ . _:b42685562 . _:b584503906 . _:b584503776 "4"^^ . _:b42685563 . . _:b584503727 . _:b584503927 . _:b584503917 . _:b584503779 "4"^^ . _:b42685572 . . _:b584503916 . _:b584503778 "4"^^ . _:b42685570 . _:b584503904 . _:b42685573 . _:b584503919 . _:b42685540 . _:b584503815 . _:b584503789 "4"^^ . _:b584504127 . _:b42685574 . _:b584503918 . _:b584503788 "4"^^ . . _:b584503913 . _:b584503791 "4"^^ . _:b42685568 . _:b584503912 . _:b584503790 "4"^^ . _:b42685569 . _:b584503915 . _:b584503785 "4"^^ . _:b42685556 . _:b42685570 . . _:b584503914 . _:b584503784 "4"^^ . _:b42685571 . . _:b584503925 . _:b584503787 "4"^^ . _:b584503886 . . _:b584503924 . _:b584503786 "4"^^ . _:b584503927 . _:b584503797 "4"^^ . _:b42685569 "search complexity, and argued that physical discontinuities or gaps within the pre-synaptic complex could further limit search complexity by segregating the Rad51-ssDNA filament into non-overlapping functional search units (Qi et\u00A0al., >>2015<<). The binding of RFS-1/RIP-1 to or within the pre-synaptic filament could define such search unit boundaries." . _:b584503926 . _:b584503796 "4"^^ . _:b584503799 "3"^^ . _:b584503921 . _:b584504029 . . _:b584503798 "3"^^ . _:b584503920 . _:b584503752 . _:b584504000 . _:b584503923 . _:b584503998 . _:b584503793 "4"^^ . _:b584503922 . _:b584503792 "4"^^ . _:b42685537 . _:b584503933 . _:b584503795 "4"^^ . _:b584503908 . . _:b584503792 . _:b584503932 . _:b584503794 "4"^^ . _:b584503805 "3"^^ . _:b584503935 . _:b42685574 . _:b584503804 "3"^^ . _:b584503934 . _:b584503807 "3"^^ . _:b42685536 . _:b584503929 . . _:b584504103 . _:b584503806 "3"^^ . _:b584503928 . _:b584503801 "3"^^ . _:b584503931 . _:b584503800 "3"^^ . _:b584503930 . _:b42685526 "HR is regulated by mediator proteins, including BRCA2, Rad54, and the family of Rad51 paralogs (San Filippo et\u00A0al., >>2008<<), which appear to act as positive regulators at different steps of the HR reaction." . _:b42685543 . . _:b584503803 "3"^^ . _:b584503941 . . _:b584504119 . _:b584503837 . . _:b584503802 "3"^^ . _:b584503940 . . . _:b584503943 . . _:b584503942 . _:b584503937 . _:b42685568 . _:b584503936 . _:b42685549 . . _:b584503939 . _:b584503974 . . _:b584503938 . _:b584503949 . _:b584503948 . . _:b584503841 . _:b584503951 . . . _:b584503950 . . _:b584504100 . _:b584503945 . _:b584503858 . _:b584503944 . . _:b584503947 . _:b42685539 "ovarian cancer (Howlett et\u00A0al., 2002; Lancaster et\u00A0al., 1996; Rahman et\u00A0al., 2007; Reid et\u00A0al., 2007; Wooster et\u00A0al., 1995; Xia et\u00A0al., 2007), biallelic germline mutations in RAD51C cause a severe form of Fanconi anemia (Vaz et\u00A0al., >>2010<<), whereas monoallelic inheritance of mutations in RAD51C and RAD51D, and RAD51B, predispose individuals to ovarian and breast cancer, respectively (Golmard et\u00A0al., 2013; Loveday et\u00A0al., 2011; Meindl et\u00A0al., 2010), demonstrating an" . _:b584503952 . . _:b584503946 . _:b42685572 . _:b42685549 . _:b584503957 . _:b584504013 . _:b584503956 . _:b584503959 . . _:b584503848 . . _:b42685561 . _:b584503958 . _:b584503953 . _:b584503965 . . . _:b584503936 . _:b584503952 . . . _:b584503955 . _:b584503986 . . _:b584503954 . _:b584503965 . _:b584504043 . _:b584503964 . _:b584503967 . _:b584504016 . _:b584503745 . _:b584503966 . . . . . _:b584503961 . . _:b584504146 . . _:b584503960 . . . _:b584503864 . _:b584503963 . _:b584503897 . . . . _:b584503962 . _:b584504111 . _:b584504099 . . _:b584503973 . . _:b584504061 . _:b584503972 . _:b42685572 . _:b42685572 . _:b584503975 . _:b584503974 . _:b584503867 . _:b584503915 . . _:b42685536 . _:b584503969 . . _:b584503968 . _:b584503894 . _:b584503971 . _:b584504151 . _:b584503970 . _:b584503810 . _:b584503817 . _:b584503934 . _:b584503757 . . _:b584503981 . . _:b584503980 . . _:b584503983 . . _:b584503982 . . _:b584503920 . . _:b584503977 . . . _:b584503976 . _:b584503746 . . . _:b584503979 . _:b584503978 . _:b584503857 . _:b584504101 . . _:b584503989 . _:b584503811 . _:b584503988 . . . _:b584503991 . _:b584503887 . . . . _:b584503990 . _:b584503985 . _:b584503987 . . _:b584503984 . _:b584504007 . _:b584504129 . . _:b584503987 . _:b584504134 . _:b584503844 . _:b584503986 . _:b584503786 . _:b584503997 . _:b584503996 . _:b42685560 "elegans BRC-2 that stabilizes RAD-51-ssDNA filaments by inhibiting RAD-51 ATP hydrolysis (Figure\u00A0S5H) (Petalcorin et\u00A0al., >>2007<<). RFS-1/RIP-1 still conferred DNaseI sensitization, confirming this assay reflects a change to\u00A0a\u00A0more \u201Copen\u201D filament conformation, rather than filament disassembly." . _:b584503796 . _:b584503999 . _:b584503766 . . . _:b584503998 . _:b584503865 . _:b584504115 . _:b584503993 . _:b42685561 . _:b584503992 . . _:b584503995 . _:b584503783 . _:b42685534 . _:b42685538 . _:b584503994 . . _:b42685538 . _:b42685538 . _:b584504005 . _:b584504004 . . _:b584504007 . _:b42685574 . . _:b584504006 . _:b584504001 . _:b584503917 . . _:b584504000 . _:b584503957 . . . _:b584504003 . . _:b584503754 . _:b584504002 . . _:b584503928 . _:b584504013 . . . _:b584504012 . _:b584504015 . _:b584503911 . _:b584504014 . _:b584504009 . _:b584503892 . . _:b584504008 . _:b584504011 . _:b42685564 . . _:b42685561 . _:b584504010 . . _:b584503780 . _:b584504021 . _:b584503940 . _:b584504020 . _:b584504023 . . _:b584504022 . _:b584504017 . . _:b584504016 . _:b584503747 . _:b584503932 . _:b584504019 . _:b584504105 . _:b584503753 . _:b584504149 . . _:b584504018 . _:b42685546 "results" . _:b584504029 . _:b42685536 . _:b584504028 . . _:b584504031 . . _:b584504030 . _:b42685561 "The Walker motifs of Rad51 paralogs are important for resistance to DNA-damaging agents in\u00A0vivo (French et\u00A0al., 2003; Gruver et\u00A0al., 2005; Wiese et\u00A0al., 2006; Yamada et\u00A0al., >>2004<<), but the\u00A0biochemical function of these motifs is unclear." . _:b584504025 . _:b584503728 . _:b42685533 "In addition, the budding and fission yeast Shu complexes (Mart\u00EDn et\u00A0al., 2006; Shor et\u00A0al., >>2005<<) contain proteins lacking detectable sequence homology to Rad51 but contain Rad51-like folds as determined by crystal structures (Sasanuma et\u00A0al., 2013; Tao et\u00A0al., 2012) or show very limited homology centered on the Walker B motif, such" . _:b584503729 . _:b584504024 . _:b584503730 . _:b584504089 . _:b584503731 . _:b584503732 . _:b584503953 . _:b584503733 . _:b584504027 . . _:b584503734 . _:b584503735 . _:b42685570 . _:b584503736 . _:b584503737 . _:b584504026 . _:b584503738 . _:b584503739 . . _:b584503740 . _:b584503764 . _:b584503741 . _:b584504037 . . _:b584503742 . _:b584503743 . _:b42685568 "This stretching may facilitate disruption of base pairing and stacking in the template duplex upon binding by the filament (Chen et\u00A0al., 2008; Danilowicz et\u00A0al., >>2014<<). The DNaseI sensitivity of the remodeled filament indicates that the ssDNA is more accessible, and it is therefore possible that the remodeling induced by Rad51 paralogs exposes the ssDNA to facilitate homology probing after filament" . _:b584503712 . _:b584503962 . _:b584503713 . _:b584504036 . _:b584503714 . _:b584503715 . _:b584503716 . _:b584503717 . _:b584504039 . _:b584503718 . _:b584503719 . _:b584504107 . _:b584503720 . _:b584503812 . _:b584503721 . _:b584504038 . _:b584504015 . _:b584503722 . _:b584503723 . _:b584503724 . _:b584503809 . _:b584503725 . _:b584504033 . _:b584503984 . _:b584503916 . _:b584503726 . _:b584503727 . . _:b584504032 . _:b584503743 . . _:b584504062 . _:b584504035 . _:b584504130 . . _:b42685573 _:b42685574 . _:b584503739 . _:b584504034 . _:b584504045 . _:b584504087 . . . _:b584504044 . . . _:b584503951 . _:b584504120 . _:b584504047 . . _:b584503878 . . _:b584503838 . _:b584504046 . _:b584504113 . _:b584504041 . _:b584503733 . _:b584503792 . _:b584503883 . . _:b584503793 . _:b584504040 . _:b584503794 . _:b584503795 . _:b584503796 . _:b584503797 . _:b584504043 . _:b584503881 . _:b584503798 . _:b584504131 . _:b584503799 . _:b584503800 . _:b584503801 . _:b584504042 . _:b584504040 . _:b584503802 . _:b584503803 . _:b584503804 . _:b584503805 . _:b584504053 . _:b584503806 . _:b584503807 . . _:b584503776 . _:b584503777 . _:b584504052 . . _:b584503778 . _:b584503779 . _:b584503780 . _:b584503781 . _:b584504055 . _:b584503782 . . _:b584503941 . _:b584503783 . . _:b584503784 . _:b584503785 . _:b584504054 . _:b584503786 . _:b584503875 . _:b584503787 . _:b584503788 . . _:b584503789 . _:b584504049 . _:b584503790 . _:b584503791 . _:b584503760 . _:b584503761 . _:b42685566 "are sensitive to both IR and nitrogen mustard, but only defective in RAD-51 focus formation in response to the latter, this likely reflects the additional function for RFS-1/RIP-1 in late stages of meiotic DSB repair (Ward et\u00A0al., >>2010<<). Furthermore, the sensitivity of rfs-1 animals to IR is relatively weak compared to deficiency in the core HR component BRC-1, whereas sensitivity of rfs-1 and brc-1 animals to crosslinking agents is virtually indistinguishable, in" . _:b584504048 . . . . _:b584503762 . _:b584503763 . _:b584503764 . _:b584503765 . _:b584504051 . _:b584503766 . . _:b584503767 . . _:b584503768 . _:b584503769 . _:b584504050 . _:b584503770 . _:b584503771 . . _:b584503772 . _:b584503890 . . _:b584503773 . _:b584504108 . _:b584504061 . _:b584503774 . _:b584503775 . . _:b584503744 . . _:b584503745 . _:b584504060 . . _:b584503746 . _:b584503747 . . _:b584503748 . _:b584503749 . _:b584504063 . _:b584503750 . _:b584503751 . _:b584503752 . _:b584503753 . _:b584504062 . . _:b584503754 . _:b584503755 . _:b584503756 . _:b584503832 . _:b584504057 . _:b584503757 . _:b584503758 . _:b584504050 . _:b584503759 . _:b584504085 . _:b584504056 . . _:b584504059 . . _:b584503740 . . . _:b584503728 . _:b584504058 . _:b584503924 . . . . _:b584503772 . _:b584503949 . . _:b42685566 . _:b584503717 . _:b584503969 . _:b584503805 . _:b584504022 . _:b584503714 . . _:b584503726 . . _:b42685532 . _:b584503954 . _:b584503901 . _:b584503989 . _:b42685555 . . _:b42685552 . _:b584504011 . . . _:b42685554 "et\u00A0al., 2006; Shor et\u00A0al., 2005), which functions in HR, and human cells encode an ATPase-sharing homology with the archael recombinase RadA, SWSAP1, required for HR, Rad51 focus formation, and DNA damage resistance (Liu et\u00A0al., 2011b)." . _:b584503849 . _:b584503798 . _:b42685527 . _:b42685545 . _:b584503833 . _:b42685573 "experimental procedures" . _:b42685547 . . _:b584504138 . . _:b584503821 . _:b584504114 . _:b42685549 "with Rad51 family proteins, but does contain a sequence resembling a Walker B motif (Figure\u00A01A), similar to the divergent yeast Rad51 paralog Psy3 that adopts a Rad51-like fold (Mart\u00EDn et\u00A0al., 2006; Sasanuma et\u00A0al., 2013; Tao et\u00A0al., >>2012<<). Since Walker B motifs mediate protein-protein interactions in human Rad51 paralog complexes (Wiese et\u00A0al., 2006), we tested if this dependency is conserved for the RFS-1/RIP-1 protein-protein interaction." . _:b584504004 . . _:b584503866 . _:b42685541 . _:b584503983 . . _:b584504126 . _:b584504039 . _:b584504143 . _:b584503995 . . _:b584503802 . _:b584503963 . . _:b584503830 . . _:b42685562 "discussion" . . . _:b42685531 "Rad51 paralogs have been identified in mammalian and avian species, which interact with one another to form two constitutive complexes, RAD51B-RAD51C-RAD51D-XRCC2 (BCDX2) and RAD51C-XRCC3 (CX3) (Masson et\u00A0al., 2001; Yonetani et\u00A0al., >>2005<<), and in budding yeast two Rad51 paralogs constitute the Rad55-Rad57 heterodimer (Sung, 1997)." . _:b584504079 . . _:b584504064 . . . _:b584503938 . _:b584503943 . _:b42685548 . _:b42685527 "For example, BRCA2 facilitates Rad51 nuclear localization (Jeyasekharan et\u00A0al., 2013; Martin et\u00A0al., >>2005<<), RPA displacement from ssDNA, and Rad51 filament nucleation (Jensen et\u00A0al., 2010; Liu et\u00A0al., 2010; Thorslund et\u00A0al., 2010), whereas Rad54 unloads Rad51 from dsDNA to promote repair DNA synthesis (Solinger et\u00A0al., 2002)." . _:b42685559 "We reasoned that a change in the structural properties of the\u00A0filaments could alter the accessibility of the ssDNA to degradation by nucleases (Zaitsev and Kowalczykowski, >>1999<<). Using a nuclease protection assay to monitor the sensitivity of pre-formed protein-DNA complexes to DNaseI, we observed that the addition of sub-stoichiometric quantities of RFS-1/RIP-1 to RAD-51 caused a dramatic and unexpected" . _:b584504056 . _:b42685544 "Shu complex form co-complexes with Rad51-ssDNA filaments (Liu et\u00A0al., 2011a; Sasanuma et\u00A0al., 2013), which in the case of Rad55-Rad57 renders the pre-synaptic complex resistant to disruption by the anti-recombinase Srs2 (Liu et\u00A0al., 2011a). However, the mechanism by which Rad51 paralogs directly stimulate the recombinase activity of Rad51 has remained enigmatic for many years (Sung, 1997)." . _:b584503931 . _:b42685565 "A requirement for Rad51 paralogs in preventing filament disruption by Srs2 at DSBs (Liu et\u00A0al., 2011a) is likely dispensable in nematodes as they lack a Srs2 ortholog." . . _:b584503914 . . . _:b584504034 . . _:b584503749 . . _:b42685553 "The existence of non-canonical Rad51 paralogs in other organisms is not unprecedented: yeast cells encode the Shu complex (Mart\u00EDn et\u00A0al., 2006; Shor et\u00A0al., >>2005<<), which functions in HR, and human cells encode an ATPase-sharing homology with the archael recombinase RadA, SWSAP1, required for HR, Rad51 focus formation, and DNA damage resistance (Liu et\u00A0al., 2011b)." . _:b584503859 . _:b584503982 . . . _:b584504023 . _:b584504112 . _:b584504113 . _:b584504114 . _:b584503840 . _:b584504115 . _:b584504116 . _:b584504020 . _:b584504117 . _:b584504118 . _:b584504119 . _:b584503948 . . _:b584504120 . _:b584504121 . _:b584504122 . _:b584504123 . . _:b584504124 . _:b584504125 . _:b584504126 . _:b584503831 . _:b584504127 . _:b584504096 . _:b584504097 . _:b584504098 . _:b584504099 . _:b584504100 . _:b584504101 . _:b584504102 . _:b584504103 . _:b584504104 . _:b584504105 . _:b584504106 . _:b584504107 . _:b584504108 . . _:b584504109 . . _:b584504110 . _:b584504111 . _:b584504080 . _:b42685574 . _:b584504017 . _:b584504081 . _:b584504082 . _:b584504083 . _:b584504021 . _:b584504084 . _:b584504085 . _:b42685572 . _:b584504086 . _:b584504087 . _:b584504044 . _:b584503961 . _:b584504088 . _:b584504091 . _:b584504082 . _:b584504089 . _:b584503879 . _:b584504090 . _:b584504091 . _:b42685531 . _:b584504092 . _:b584504093 . _:b584504094 . . _:b584504095 . _:b584504064 . _:b584503813 . . _:b584504065 . _:b584504066 . . _:b584504067 . . _:b584504068 . _:b584504069 . _:b584504070 . _:b584504071 . _:b584504072 . . _:b584504073 . _:b584504074 . _:b584503795 . _:b584504075 . _:b584504076 . _:b584503828 . . _:b42685541 "The budding yeast Rad55-Rad57 complex (Sung, >>1997<<) and a sub-complex of the human BCDX2 complex, RAD51B-RAD51C (Sigurdsson et\u00A0al., 2001), have been purified as heterodimers and despite lacking intrinsic recombinase activity they stimulate strand exchange by Rad51 in\u00A0vitro." . _:b584504141 . _:b584504077 . _:b584504078 . . _:b584504079 . . _:b584503813 "3"^^ . _:b584503812 "3"^^ . . _:b584504160 . _:b584503815 "3"^^ . _:b584503814 "3"^^ . _:b584503809 "3"^^ . _:b584503829 . _:b584503808 "3"^^ . _:b584504144 . _:b584503850 . _:b584504145 . _:b584504146 . _:b584503811 "3"^^ . _:b584504147 . _:b584504148 . _:b584504149 . . _:b584504150 . _:b584504139 . _:b584503810 "3"^^ . _:b584504151 . _:b584504152 . . _:b584504153 . _:b584504154 . _:b584503925 . _:b584503821 "3"^^ . _:b584504155 . . _:b584504156 . _:b584504157 . _:b584503818 . _:b584504158 . _:b584503820 "3"^^ . _:b584503884 . _:b584504159 . _:b584504128 . _:b584504129 . . _:b584504130 . . _:b584503779 . _:b584503823 "3"^^ . _:b584504131 . . _:b42685537 . . . _:b584504132 . . _:b584504133 . _:b584504134 . _:b584503822 "3"^^ . _:b42685570 "process, facilitated by transient contacts with heterologous dsDNA to enhance the probability of locating homologous sequences (Forget and Kowalczykowski, 2012; Gonda and Radding, 1983, 1986; Honigberg et\u00A0al., 1986; Tsang et\u00A0al., >>1985<<). The increased flexibility of the remodeled filament induced by RFS-1/RIP-1 in 3D space could aid such a 3D homology search mechanism." . _:b584504135 . _:b584503816 . _:b584504136 . . _:b584504137 . _:b584504138 . _:b584503817 "3"^^ . _:b584504057 . _:b584503944 . _:b584504139 . _:b584504140 . . _:b584504141 . _:b584504142 . _:b584503816 "3"^^ . _:b584503909 . _:b584504143 . . _:b584504068 . . _:b584503819 "3"^^ . . _:b584503818 "3"^^ . _:b584503732 . _:b584503958 . _:b584503829 "3"^^ . _:b584503828 "3"^^ . _:b584503955 . _:b584503831 "3"^^ . _:b584504054 . _:b584503830 "3"^^ . _:b584504035 . _:b584503825 "3"^^ . . . _:b584503900 . _:b584503824 "3"^^ . . _:b584503736 . _:b584503827 "3"^^ . _:b584503826 "3"^^ . . _:b584504069 . . _:b584503837 "3"^^ . . _:b584504068 . _:b584503773 . _:b584503836 "3"^^ . _:b584504071 . _:b584503768 . _:b584504058 . _:b584503839 "3"^^ . _:b584504070 . _:b584503838 "3"^^ . _:b584504065 . _:b584503833 "3"^^ . _:b584503723 . _:b584504064 . _:b584503994 . _:b584503832 "3"^^ . _:b584503719 . _:b584504067 . . _:b584503835 "3"^^ . _:b584504066 . _:b584503834 "3"^^ . _:b584504077 . _:b584503845 "3"^^ . _:b584504080 . . _:b584504076 . . _:b584503844 "3"^^ . _:b584503718 . _:b584504079 . . _:b584504063 . _:b584504053 . . _:b584503847 "3"^^ . _:b584504078 . . _:b584503846 "3"^^ . _:b584504073 . _:b584503841 "3"^^ . _:b584504072 . . _:b42685536 . _:b584503840 "3"^^ . . _:b584504075 . _:b584503843 "3"^^ . _:b584504073 . _:b584504074 . _:b42685550 . _:b584503842 "3"^^ . _:b584504085 . _:b42685571 "Our observations, together with previously reported findings, suggest that Rad51 paralogs perform two distinct functions to promote HR: they protect Rad51-ssDNA filaments against disruption by antirecombinases (Liu et\u00A0al., 2011a) and directly stimulate the intrinsic recombinase activity of Rad51 by remodeling pre-synaptic filaments to an active, \u201Copen,\u201D flexible, and stable conformation primed for homology search and strand invasion." . _:b584503853 "3"^^ . _:b584504084 . _:b584503785 . . _:b584503852 "3"^^ . . _:b584504087 . . _:b584503855 "3"^^ . _:b584504086 . . _:b584503854 "3"^^ . _:b584504081 . _:b42685542 . . . . . _:b584503849 "3"^^ . _:b584504080 . _:b584503825 . _:b584503848 "3"^^ . _:b42685570 . _:b584503856 . _:b584504083 . _:b584503857 . _:b584503858 . _:b584503859 . _:b584503851 "3"^^ . . _:b584503744 . _:b584504082 . _:b584503860 . . _:b584503991 . _:b584503861 . _:b584503862 . _:b42685539 . _:b584503863 . _:b584503850 "3"^^ . _:b584503864 . _:b584504093 . _:b584503865 . _:b584503866 . _:b584503867 . _:b42685540 . _:b584503861 "3"^^ . . _:b584504092 . _:b584503868 . _:b584503869 . _:b584503870 . _:b584503871 . _:b584503860 "3"^^ . _:b584503840 . _:b584504095 . . _:b584503841 . . _:b584503842 . _:b584503843 . _:b584503863 "3"^^ . . _:b584504094 . _:b584503844 . . _:b584503845 . _:b584503846 . _:b584503847 . _:b584503862 "3"^^ . _:b584503776 . _:b584503848 . _:b584504089 . _:b584503770 . _:b584503849 . . _:b584503850 . _:b584503851 . _:b584503857 "3"^^ . . _:b584504088 . _:b584503742 . _:b584503852 . . _:b584503853 . . _:b584503854 . _:b584503855 . _:b584503889 . _:b584503856 "3"^^ . . _:b42685545 "However, the mechanism by which Rad51 paralogs directly stimulate the recombinase activity of Rad51 has remained enigmatic for many years (Sung, >>1997<<). Additionally, whether the Rad51 paralogs confer any intrinsic stabilization or alteration in the structural properties of the pre-synaptic complex is unknown, as is the mechanistic importance of their conserved Walker motifs." . _:b584504091 . _:b584503824 . _:b584503825 . . _:b584503826 . _:b584503827 . _:b42685558 . _:b584503859 "3"^^ . _:b584503828 . _:b584504090 . _:b584503731 . . _:b584503829 . _:b584503830 . . _:b584503831 . _:b584503858 "3"^^ . _:b584503832 . _:b584504101 . _:b584503833 . _:b584503834 . _:b584504055 . _:b584503835 . _:b584503869 "3"^^ . _:b584503836 . _:b584504100 . _:b584503837 . _:b584503838 . _:b584503839 . _:b584503868 "3"^^ . _:b584503808 . _:b584504103 . _:b584503809 . . _:b584503810 . . _:b584503871 "3"^^ . _:b584503811 . _:b584503812 . _:b584504102 . _:b584503813 . _:b42685532 "another to form two constitutive complexes, RAD51B-RAD51C-RAD51D-XRCC2 (BCDX2) and RAD51C-XRCC3 (CX3) (Masson et\u00A0al., 2001; Yonetani et\u00A0al., 2005), and in budding yeast two Rad51 paralogs constitute the Rad55-Rad57 heterodimer (Sung, >>1997<<). In addition, the budding and fission yeast Shu complexes (Mart\u00EDn et\u00A0al., 2006; Shor et\u00A0al., 2005) contain proteins lacking detectable sequence homology to Rad51 but contain Rad51-like folds as determined by crystal structures (Sasanuma" . _:b584503814 . _:b584503815 . _:b584503870 "3"^^ . _:b584503860 . _:b584503816 . _:b584504097 . _:b584503817 . _:b584503818 . _:b42685561 . _:b584503819 . _:b42685570 . _:b584503877 . _:b584503865 "3"^^ . _:b584503820 . _:b584504096 . _:b584503821 . _:b584503822 . _:b584503823 . _:b584503864 "3"^^ . _:b584503920 . _:b584504099 . _:b584503921 . _:b584503922 . _:b584503923 . _:b584503867 "3"^^ . . _:b584504098 . _:b584504086 . _:b584503924 . _:b584503925 . _:b584503926 . _:b584503927 . _:b584503866 "3"^^ . . _:b584504109 . _:b584503928 . _:b584503929 . _:b584503919 . _:b584503930 . _:b584504009 . _:b584503931 . _:b42685574 "Yeast two-hybrid and nematode genetic analysis were performed as previously described (Boulton et\u00A0al., 2002; Ward et\u00A0al., 2007, >>2010<<). Full materials and methods, including details of stopped-flow data analysis and smFRET, are available in the Supplemental Experimental Procedures." . _:b584503877 "3"^^ . . _:b584504108 . _:b584503932 . "PMC0" . _:b584503933 . _:b584503934 . _:b584503935 . _:b584503876 "3"^^ . _:b584503904 . _:b584504111 . _:b584503905 . . _:b584503906 . _:b42685546 _:b42685548 . _:b42685546 _:b42685549 . _:b584503879 "3"^^ . _:b584503907 . _:b42685546 _:b42685550 . _:b584504110 . _:b584503872 . _:b584503908 . _:b42685546 _:b42685551 . _:b584503909 . _:b584503910 . _:b584503911 . _:b584503878 "3"^^ . _:b584503912 . _:b584504105 . . _:b42685546 _:b42685547 . _:b584503996 . _:b584503913 . _:b584503914 . _:b584503915 . _:b584504083 . _:b584503873 "3"^^ . _:b584503916 . _:b584504104 . . _:b584504088 . _:b584503917 . _:b584503918 . _:b584503919 . _:b584503872 "3"^^ . . _:b584504107 . _:b584503888 . _:b584503889 . _:b584503890 . _:b584503891 . _:b584503875 "3"^^ . _:b584503892 . _:b584504106 . _:b584503893 . _:b42685546 _:b42685560 . _:b584503894 . _:b42685546 _:b42685561 . _:b584503874 "3"^^ . _:b584503895 . _:b584503896 . _:b584504117 . _:b584503897 . _:b42685546 _:b42685556 . _:b584503898 . _:b42685546 _:b42685557 . _:b584503885 "3"^^ . _:b584503871 . _:b584503899 . . _:b584504116 . _:b584503900 . _:b42685546 _:b42685558 . . . _:b584503901 . _:b42685546 _:b42685559 . _:b42685546 _:b42685552 . _:b584503902 . _:b42685546 _:b42685553 . _:b584503884 "3"^^ . _:b584503903 . _:b584503750 . _:b584504119 . _:b584503872 . _:b42685546 _:b42685554 . _:b42685546 _:b42685555 . . _:b584503873 . _:b584503874 . _:b584503875 . _:b584503887 "3"^^ . _:b584503876 . _:b584504118 . _:b584503877 . _:b584503878 . _:b584503879 . _:b584503886 "3"^^ . _:b584503880 . _:b584504113 . . _:b584503881 . _:b584503845 . _:b584503882 . _:b584503883 . _:b584504072 . _:b584503881 "3"^^ . _:b584503884 . _:b584504125 . _:b584504112 . _:b584503885 . _:b584503822 . _:b584503886 . . _:b584503880 "3"^^ . _:b584503887 . . _:b584504115 . _:b584503984 . . . _:b584503985 . . . _:b584503986 . . _:b584503883 "3"^^ . _:b584503987 . _:b584503988 . _:b584504114 . . _:b584503989 . _:b584503990 . _:b584504144 . _:b584503991 . _:b584503882 "3"^^ . _:b584503992 . _:b584504125 . _:b584503993 . _:b584503994 . _:b584503893 "2"^^ . _:b584503995 . _:b584503836 . _:b584504124 . _:b584503996 . . _:b584504122 . _:b584503997 . _:b584503717 . . _:b584503892 "2"^^ . _:b584503998 . _:b584503999 . _:b584503855 . _:b584503968 . _:b584504127 . _:b584503716 . _:b584503992 . _:b584503969 . _:b584503970 . _:b584503895 "2"^^ . _:b584503971 . _:b584503972 . _:b584504126 . _:b584503806 . _:b584503973 . _:b584503719 . . _:b584503894 "2"^^ . _:b584503974 . . _:b584503975 . _:b584503715 . _:b584503725 . _:b584504121 . _:b584503898 . _:b584503976 . _:b584503718 . _:b584503977 . _:b584503978 . _:b584503979 . _:b584503889 "3"^^ . _:b584503980 . _:b584504120 . _:b584503713 . _:b584503981 . _:b584503982 . _:b584503983 . _:b584503888 "3"^^ . _:b584503952 . _:b584504123 . _:b584503953 . _:b584503977 . . _:b584503712 . _:b584503954 . _:b42685562 _:b42685564 . _:b584503955 . _:b584503891 "3"^^ . _:b42685562 _:b42685565 . . _:b584503956 . _:b584504122 . _:b584504095 . _:b42685562 _:b42685566 . _:b584503957 . _:b584504078 . . _:b42685562 _:b42685567 . _:b584503715 . _:b584503958 . _:b584503959 . _:b584503890 "3"^^ . _:b584503960 . _:b584504133 . _:b584503961 . _:b584504117 . _:b42685562 _:b42685563 . _:b584503714 . _:b584503962 . _:b584503901 "2"^^ . _:b584503963 . _:b584503774 . . _:b584503964 . _:b584504132 . _:b584503965 . _:b584503725 . _:b584503966 . _:b584504051 . _:b584503900 "2"^^ . _:b584503967 . _:b584503936 . _:b584504135 . _:b584503937 . _:b584503724 . _:b584503938 . _:b584504156 . _:b584503903 "2"^^ . . _:b584503939 . . _:b584503940 . _:b584504134 . _:b584503941 . _:b584503727 . _:b584503942 . _:b584503902 "2"^^ . . _:b584503943 . _:b584503944 . _:b584504129 . . _:b584503945 . _:b584504136 . _:b584503726 . . . _:b584503946 . _:b584503897 "2"^^ . _:b42685562 _:b42685572 . . _:b584503947 . _:b584504142 . . . _:b584503948 . _:b584504128 . _:b584503949 . _:b584503771 . _:b584503721 . _:b584503950 . _:b584503896 "2"^^ . _:b42685562 _:b42685568 . _:b584503951 . _:b42685562 _:b42685569 . . _:b584504048 . _:b584504131 . _:b42685562 _:b42685570 . _:b584504049 . _:b42685562 _:b42685571 . _:b584503720 . _:b584504050 . _:b584503899 "2"^^ . . _:b584504051 . _:b584504052 . _:b584504130 . . _:b584504053 . _:b584503723 . _:b584504054 . _:b584503898 "2"^^ . _:b584504055 . _:b584504056 . _:b584504141 . _:b584504057 . _:b584503722 . . _:b584504058 . _:b42685570 . _:b584503909 "2"^^ . . _:b584504059 . _:b584504060 . _:b584504140 . _:b584504061 . _:b584503733 . _:b584504062 . _:b584504065 . _:b584503908 "2"^^ . _:b584504063 . _:b584504032 . _:b584504143 . _:b584504033 . _:b584503732 . _:b584504034 . _:b584503911 "2"^^ . . _:b584503762 . _:b584504035 . _:b584504036 . _:b42685529 "Martin et\u00A0al., 2005), RPA displacement from ssDNA, and Rad51 filament nucleation (Jensen et\u00A0al., 2010; Liu et\u00A0al., 2010; Thorslund et\u00A0al., 2010), whereas Rad54 unloads Rad51 from dsDNA to promote repair DNA synthesis (Solinger et\u00A0al., >>2002<<). However, the molecular mechanism underlying the stimulation of HR by the Rad51 paralogs has remained elusive." . _:b584504142 . . _:b584504037 . _:b584504067 . _:b584503735 . _:b584504038 . _:b584503929 . _:b584503910 "2"^^ . _:b584504039 . . _:b584504040 . _:b584504137 . _:b584503851 . _:b584504041 . _:b584503734 . _:b584504042 . _:b584503905 "2"^^ . _:b584504043 . _:b584504044 . _:b584504136 . _:b584504045 . _:b584504032 . _:b584503729 . _:b584504046 . _:b584503904 "2"^^ . . _:b584504047 . _:b584504016 . _:b584504139 . . _:b584504017 . _:b584503728 . _:b584503808 . _:b584504018 . _:b584503907 "2"^^ . _:b584504019 . _:b584504020 . _:b584504138 . _:b584504021 . _:b584503731 . _:b584504022 . _:b584503906 "2"^^ . . _:b584504023 . _:b584504024 . _:b584504149 . . _:b584504025 . _:b584503730 . _:b584504026 . _:b584503917 "2"^^ . _:b584504027 . . _:b584504028 . _:b584504148 . _:b584504029 . _:b584503741 . _:b584504030 . _:b584503916 "2"^^ . _:b584503782 . _:b584504031 . _:b584504000 . _:b584504151 . . _:b584504001 . . _:b584503740 . _:b584504002 . _:b584503919 "2"^^ . _:b584504003 . _:b584504049 . _:b584504004 . _:b584504150 . . . _:b584504005 . _:b584503743 . _:b584504006 . _:b584503918 "2"^^ . _:b584503820 . _:b584504007 . _:b584504008 . _:b584504145 . "10.1016%2Fj.cell.2015.06.015" . _:b584504009 . _:b584503742 . _:b584504010 . _:b584503973 . _:b584503913 "2"^^ . _:b584504011 . _:b584504012 . _:b584504144 . _:b584504013 . _:b42685538 . _:b584503737 . _:b584504014 . _:b584503912 "2"^^ . _:b584504015 . _:b42685558 . . _:b584504147 . _:b584503736 . _:b42685524 _:b42685540 . _:b584503915 "2"^^ . _:b42685524 _:b42685541 . _:b42685524 _:b42685542 . _:b584504146 . _:b584503739 . _:b42685524 _:b42685543 . . _:b584503914 "2"^^ . _:b42685524 _:b42685536 . _:b42685524 _:b42685537 . _:b42685524 _:b42685538 . _:b584504157 . _:b584503738 . _:b42685524 _:b42685539 . _:b584503925 "2"^^ . . _:b584504156 . _:b584503749 . _:b42685524 _:b42685544 . _:b584503924 "2"^^ . _:b42685524 _:b42685545 . _:b584504159 . _:b584503748 . . _:b584503927 "2"^^ . . _:b42685524 _:b42685525 . _:b42685524 _:b42685526 . _:b584504158 . _:b584503751 . _:b42685524 _:b42685527 . _:b584503926 "2"^^ . _:b584504153 . _:b584503750 . _:b584503729 . . _:b584503921 "2"^^ . _:b42685524 _:b42685532 . _:b42685524 _:b42685533 . _:b584504121 . . _:b584504152 . _:b42685524 _:b42685534 . . _:b42685524 _:b42685535 . _:b584503745 . _:b584503751 . _:b42685524 _:b42685528 . _:b584503975 . _:b584503920 "2"^^ . _:b584503777 . _:b42685524 _:b42685529 . . _:b584504155 . _:b42685524 _:b42685530 . _:b584503744 . _:b42685524 _:b42685531 . _:b584503923 "2"^^ . . _:b584504154 . _:b584503747 . . _:b584503922 "2"^^ . . _:b584503746 . _:b584503933 "2"^^ . _:b584504137 . . . _:b584504140 . _:b584503801 . _:b584503757 . . _:b584503932 "2"^^ . _:b584504158 . . _:b584503756 . _:b584503935 "2"^^ . _:b584503759 . _:b584503835 . _:b584503934 "2"^^ . . _:b584503721 . . _:b584503758 . _:b584503846 . _:b584503929 "2"^^ . _:b584504160 . _:b584503753 . _:b584503928 "2"^^ . . . _:b584503752 . _:b584503931 "2"^^ . _:b584503755 . _:b584503876 . _:b584503930 "2"^^ . _:b584503756 . . _:b584503754 . . _:b584503941 "2"^^ . _:b584503765 . _:b584503940 "2"^^ . _:b584503947 . _:b584503764 . _:b584503943 "2"^^ . _:b584504132 . _:b584503888 . _:b584503767 . _:b584503942 "2"^^ . _:b584503766 . _:b584503937 "2"^^ . _:b42685542 "The budding yeast Rad55-Rad57 complex (Sung, 1997) and a sub-complex of the human BCDX2 complex, RAD51B-RAD51C (Sigurdsson et\u00A0al., >>2001<<), have been purified as heterodimers and despite lacking intrinsic recombinase activity they stimulate strand exchange by Rad51 in\u00A0vitro." . _:b584503870 . _:b584503761 . _:b584503936 "2"^^ . . _:b584503734 . . _:b584504028 . _:b584503760 . _:b584503939 "2"^^ . _:b584503970 . _:b584503763 . _:b584504052 . _:b584503938 "2"^^ . _:b584503762 . _:b584504037 . _:b584503949 "2"^^ . . _:b584503773 . . _:b584503948 "2"^^ . _:b584503905 . . . _:b584503772 . . _:b584503951 "2"^^ . _:b584503775 . _:b584503950 "2"^^ . _:b584503774 . _:b584503945 "2"^^ . _:b584503769 . _:b584503944 "2"^^ . . . _:b584504160 . _:b584503768 . _:b584503947 "2"^^ . _:b42685527 . _:b584503771 . _:b584503946 "2"^^ . _:b584503770 . _:b584503957 "2"^^ . _:b584504118 . _:b584503839 . _:b584504076 . _:b584503724 . _:b584503781 . _:b584503956 "2"^^ . _:b584504094 . _:b584503869 . _:b584503780 . _:b584503959 "2"^^ . _:b584503783 . _:b584503958 "2"^^ . . _:b584503782 . _:b584503953 "2"^^ . _:b584503777 . _:b584503952 "2"^^ . _:b42685548 "Since Rad51 paralogs in other organisms function as complexes, we reasoned that RFS-1 may require a binding partner for optimal function. Using a yeast two-hybrid screen (Boulton et\u00A0al., >>2002<<), we identified an orphan protein encoded by R01H10.5 (Uniprot ID code Q21621) that we named RIP-1 (RFS-1 interacting protein) (Figure\u00A01A), which interacted with RFS-1 by yeast two-hybrid (Figure\u00A01B), glutathione S-transferase (GST)" . . . _:b584503776 . _:b584503955 "2"^^ . _:b584503779 . _:b584503954 "2"^^ . _:b584504092 . _:b584504019 . _:b584503778 . _:b584503965 "2"^^ . . _:b584503789 . . . _:b584503964 "2"^^ . _:b584503935 . _:b584503788 . _:b584503967 "2"^^ . . . _:b584503791 . . _:b584503966 "2"^^ . . _:b584503769 . _:b584503790 . _:b584503961 "2"^^ . . _:b584503785 . . _:b584503960 "2"^^ . . _:b584503784 . _:b584503963 "2"^^ . _:b584503827 . _:b584503787 . _:b584504148 . _:b584503856 . _:b584503962 "2"^^ . . _:b584503786 . . _:b584503973 "2"^^ . _:b584503824 . . _:b584503797 . _:b584503972 "2"^^ . _:b42685538 "Like BRCA2 and PALB2, which are mutated in Fanconi anemia and breast and ovarian cancer (Howlett et\u00A0al., 2002; Lancaster et\u00A0al., 1996; Rahman et\u00A0al., 2007; Reid et\u00A0al., 2007; Wooster et\u00A0al., 1995; Xia et\u00A0al., >>2007<<), biallelic germline mutations in RAD51C cause a severe form of Fanconi anemia (Vaz et\u00A0al., 2010), whereas monoallelic inheritance of mutations in RAD51C and RAD51D, and RAD51B, predispose individuals to ovarian and breast cancer," . . _:b584503796 . _:b584503975 "2"^^ . _:b584503799 . _:b584503974 "2"^^ . _:b584504071 . _:b584503798 . _:b584503969 "2"^^ . _:b584504077 . . _:b584503793 . _:b584503968 "2"^^ . _:b42685544 . . _:b584503792 . _:b584503891 . _:b584503971 "2"^^ . . _:b584503795 . _:b584503981 . . _:b584503970 "2"^^ . _:b584503794 . _:b584503981 "2"^^ . _:b584504110 . _:b584503823 . _:b584503852 . _:b584503805 . _:b584503980 "2"^^ . . _:b584503804 . _:b584503983 "2"^^ . _:b584503800 . . . _:b584503807 . _:b584503899 . _:b42685525 "of Rad51 from double-stranded DNA (dsDNA) permits the initiation of repair DNA synthesis and the resulting joint molecules are processed by various enzymes to complete the repair reaction (Chapman et\u00A0al., 2012; San Filippo et\u00A0al., >>2008<<)." . _:b584503982 "2"^^ . . _:b584503806 . . _:b584503977 "2"^^ . _:b584503868 . _:b584503801 . _:b584503976 "2"^^ . _:b584504133 . _:b584503800 . _:b584504048 . _:b584503979 "2"^^ . . _:b42685543 . . _:b584503803 . _:b584503978 "2"^^ . _:b584504128 . . _:b584504012 . _:b584503802 . _:b584503763 . _:b584503989 "2"^^ . _:b584503988 "2"^^ . _:b584503972 . _:b584503991 "2"^^ . . _:b584503990 "2"^^ . _:b584503976 . _:b584503985 "2"^^ . _:b584503984 "2"^^ . . _:b42685533 . _:b584503987 "2"^^ . _:b42685525 . . . _:b584504135 . _:b584503986 "2"^^ . . _:b584503854 . . . _:b584503997 "2"^^ . _:b42685573 . _:b42685565 . . _:b584503996 "2"^^ . . . _:b584503999 "2"^^ . _:b42685559 . _:b584503930 . . _:b584503998 "2"^^ . _:b584503748 . . . _:b584503993 "2"^^ . . _:b42685567 "HR component BRC-1, whereas sensitivity of rfs-1 and brc-1 animals to crosslinking agents is virtually indistinguishable, in support of the greater specificity of RFS-1/RIP-1 toward stalled replication fork substrates (Ward et\u00A0al., >>2007<<)." . . _:b584503992 "2"^^ . _:b584503995 "2"^^ . _:b584503994 "2"^^ . _:b584504002 . _:b584503788 . . _:b584503819 . _:b584504005 "2"^^ . . _:b584504014 . _:b584503775 . _:b584504004 "2"^^ . _:b42685546 . _:b42685556 . _:b42685537 . . _:b584504007 "2"^^ . _:b584503918 . _:b584503960 .