_:b18369179 . _:b18369116 "Since MCF7 cells do not express caspase-3 [>>42<<], for this set of experiments we monitored apoptosis solely by Hoechst staining." . . _:b18369129 . _:b18369103 "Total RNA extraction, cDNA synthesis and Q-PCR were performed as described previously [>>67<<]. Amplifications of the GAPDH and RPL13 genes were performed to ensure unambiguous comparisons between cDNAs from different samples. Primers and annealing temperatures for all genes are indicated in Table S1. For each gene, the values were" . . . _:b504332959 . _:b504332923 . _:b504332951 . _:b18369061 _:b18369088 . _:b18369061 _:b18369089 . _:b18369061 _:b18369090 . _:b18369061 _:b18369091 . _:b18369061 _:b18369092 . _:b18369177 "In agreement with our results, it has been shown that in cancers where TEAD1 is overexpressed, such as in prostate cancers [33], Livin is also up-regulated [64], [>>65<<]. However, interestingly, a modest but significant, increase in Livin is also observed in other types of cancers where TEAD1 is down-regulated, such as breast, renal or bladder cancers (www.oncomine.org). The data presented in our study," . . _:b18369061 _:b18369093 . _:b18369061 _:b18369094 . _:b18369061 _:b18369095 . _:b18369061 _:b18369096 . _:b18369061 _:b18369097 . _:b18369061 _:b18369098 . _:b504332925 . _:b18369061 _:b18369099 . _:b18369061 _:b18369100 . _:b18369061 _:b18369101 . _:b504332946 . _:b18369061 _:b18369072 . _:b18369061 _:b18369073 . _:b18369094 . _:b18369061 _:b18369074 . _:b504332927 . _:b18369061 _:b18369075 . _:b18369061 _:b18369076 . . _:b18369061 _:b18369077 . _:b18369061 _:b18369078 . _:b18369061 _:b18369079 . _:b18369130 . _:b18369101 "Interestingly, we show that overexpression of a mutated form of TEAD1 (Y421H) implicated in Sveinsson's chorioretinal atrophy that strongly reduces interaction of the mutant with YAP as well as its activation [38], [>>39<<] can induce Livin expression and protect cells from induced apoptosis, suggesting that YAP is not the cofactor involved in Livin repression." . _:b18369061 _:b18369080 . _:b18369061 _:b18369081 . _:b18369061 _:b18369082 . _:b18369095 . _:b18369061 _:b18369083 . _:b18369061 _:b18369084 . _:b18369061 _:b18369085 . _:b18369061 _:b18369086 . _:b18369064 "TEAD1 belongs to the family of conserved eukaryotic transcription factors (TEAD proteins), characterized by the TEA/ATTS DNA binding domain [1], [2], [>>3<<]. There are four closely related Tead genes (Tead1 to Tead4) in mammals [4], [5] and one, scalloped (sd), in Drosophila [6]." . _:b18369061 _:b18369087 . _:b18369171 "4C,D) and published data [>>62<<], [63] suggest that, in HeLa cells, YAP overexpression or the Hippo signaling pathway does not play a role in apoptosis regulation." . _:b18369061 _:b18369062 . _:b18369061 _:b18369063 . _:b18369061 _:b18369064 . _:b504332905 . . . _:b18369078 . _:b18369061 _:b18369065 . _:b18369061 _:b18369066 . _:b18369061 _:b18369067 . _:b18369061 _:b18369068 . _:b18369175 "In agreement with our results, it has been shown that in cancers where TEAD1 is overexpressed, such as in prostate cancers [>>33<<], Livin is also up-regulated [64], [65]." . _:b18369061 _:b18369069 . _:b18369061 _:b18369070 . _:b18369061 _:b18369071 . _:b504332918 . _:b18369081 . _:b18369131 . _:b18369099 . _:b18369146 "This mutation causes the human genetic disease known as Sveinsson's chorioretinal atrophy [38], and strongly reduces both YAP/TEAD1 interaction and activity [>>55<<]. Contrary to wild-type TEAD1, TEAD1-H was unable to activate the expression of the TEAD1 responsive reporter when transfected with YAP, confirming that the interaction with YAP is severely impaired (Fig. S2A). HeLa cells transfected with" . _:b504332893 . _:b504332944 . _:b18369105 "Alcaide) [>>69<<] were maintained in RPMI 1640 medium with 5% foetal calf serum for HeLa and 10% for MCF7 and BUA cells, 100 U/ml penicillin, and 100 \u00B5g/ml streptomycin at 37\u00B0C in 5% CO2." . . _:b18369074 "Yki is the Drosophila ortholog of mammalian YAP (Yes-Associated Protein) which in vitro and in vivo is a well characterized cofactor of the mammalian TEAD proteins [13], [14], [>>15<<], [16]. Both Yki and YAP, are the effectors of the Hippo tumour suppressor pathway that restricts organ growth by keeping in check cell proliferation and promoting apoptosis in Drosophila and in mammals [17], [18]. The regulation of" . . _:b504332895 . _:b504332894 . _:b18369169 . . _:b18369154 "highly limiting, possibly cell-specific, titratable transcriptional co-factor(s) has been deduced from transfection analyses where activation of a cognate reporter is severely reduced upon TEAD1 overexpression [1], [9], [56], [57], [>>58<<] (Fig. S2A)." . . _:b504332901 . _:b18369104 "For qualitative RT-PCR of Livin, primers and experimental conditions were according to [>>45<<]." . _:b504332900 . _:b504332948 . _:b18369118 "This is in contrast with previously published data in different cellular models [>>15<<], [16], indicating that the loss of function for TEAD1 sensitizes cells to apoptosis." . _:b504332903 . _:b18369136 . _:b18369132 "To gain insight into the mechanism underlying TEAD1 dependent Livin up-regulation, HeLa cells were co-transfected with a luciferase reporter construct containing the upstream promoter of Livin (ML-IAP Pro4) [>>51<<] and with the TEAD1-expressing plasmid (Fig. 3A), which resulted in a 3.4-fold reporter activation, compared to the control (Fig." . _:b504332902 . _:b504332912 . _:b18369151 "existence of such highly limiting, possibly cell-specific, titratable transcriptional co-factor(s) has been deduced from transfection analyses where activation of a cognate reporter is severely reduced upon TEAD1 overexpression [1], [>>9<<], [56], [57], [58] (Fig. S2A)." . _:b504332897 . _:b18369178 "Furthermore, TEAD1 directly regulates the transcription of Mesothelin (MSLN) that is highly expressed in several cancers and is a good candidate for a diagnostic marker [>>34<<]. Interestingly, none of the expression patterns of the known TEAD1 cofactors such as YAP matched the expression of MSLN in pancreatic cancers or in other cell line models used here." . _:b18369163 . _:b504332896 . _:b504332899 . _:b18369130 "2G and 2H), consistent with previously published data [46], [48], [>>49<<], [50]. Importantly Livin RNAi also completely abolished the resistance to STS-induced apoptosis conferred by TEAD1 overexpression (Fig." . . _:b504332898 . _:b504332909 . _:b504332908 . _:b18369139 "S2B) while the TEAD-ENRD fusion protein strongly repressed that same reporter [>>53<<] (Fig. S2A)." . _:b504332911 . _:b18369114 "Importantly, increased apoptosis was observed in the ectoderm of Tead1; Tead2 homozygous mutant mice [>>16<<] but the bona fide target genes relevant to cell survival had so far not been identified." . _:b504332910 . _:b18369104 . . _:b18369086 "This is achieved in Drosophila through the transcriptional induction of target genes including Cyclin E, dE2F1 [10], the bantam microRNA [22], [23], dmyc [>>24<<], [25] and the Drosophila inhibitor of apoptosis protein 1 (Diap1) [17]." . _:b504332905 . _:b504332904 . . _:b504332907 . _:b504332957 . . _:b18369081 "Conversely,Yki overexpression promotes organ growth by stimulating cell proliferation and preventing apoptosis [>>19<<], [21]. This is achieved in Drosophila through the transcriptional induction of target genes including Cyclin E, dE2F1 [10], the bantam microRNA [22], [23], dmyc [24], [25] and the Drosophila inhibitor of apoptosis protein 1 (Diap1) [17]." . _:b504332906 . _:b504332901 . _:b18369162 "Its disruption in mice leads to heart defects and embryonic lethality between embryonic days 11 and 12 [60] while its overexpression in the mouse heart can induce age-dependent dysfunction [>>61<<]. Moreover, recent data support an anti-apoptotic role for TEAD proteins during mouse development, based on Tead1 and/or Tead2 loss of function experiments [15], [16]. However, the effect of a gain of function for wild-type TEAD proteins" . . _:b504332898 . _:b504332917 . _:b18369119 "This is in contrast with previously published data in different cellular models [15], [>>16<<], indicating that the loss of function for TEAD1 sensitizes cells to apoptosis." . . _:b504332916 . . _:b504332919 . _:b18369141 . _:b504332939 . _:b504332918 . _:b18369140 . _:b18369169 "S1D) while they are able to interact with YAP [>>14<<]. By using deleted forms of TEAD1, we showed that both the N-terminus and C-terminus domains of TEAD1 are required for Livin up-regulation." . _:b504332913 . _:b504332912 . _:b18369121 "This hypothesis is consistent with data previously observed in cultured cells [>>9<<]." . . _:b504332915 . _:b504332914 . _:b504332912 . _:b18369092 "define a highly conserved family of intracellular proteins, the Inhibitor of Apoptosis Proteins (IAP) that suppress apoptosis induced by a variety of stimuli by binding specific intracellular proteases, primarily caspases 3, 7 and 9 [>>28<<], [29], [30]. In humans, eight family members have been identified (NAIP, c-IAP1, c-IAP2, XIAP, Survivin, Apollon, Livin and ILP2) [31], and only two in Drosophila (Diap1/2) [32]." . _:b504332913 . _:b504332914 . _:b504332925 . _:b504332915 . _:b504332916 . _:b504332917 . _:b504332918 . _:b504332924 . _:b504332919 . _:b504332920 . _:b504332893 . _:b504332921 . _:b18369093 . _:b504332922 . _:b504332927 . _:b504332923 . _:b504332924 . _:b504332925 . _:b18369168 "The anti-apoptotic features of Livin are well known [45], [65], [>>66<<]. Our Livin RNAi experiments not only confirm these previously published data, but also demonstrate that Livin up-regulation is required for TEAD1-induced apoptotic protection." . . _:b504332926 . _:b504332926 . _:b504332927 . . _:b504332896 . _:b504332897 . _:b504332898 . . _:b18369124 "In vivo, the two known spliced isoforms of Livin mRNA present different antiapoptotic properties [>>45<<], [46]. Using Jurkat cells as a model, Ashhab et al. (2001) showed that Livin \u03B1but not \u03B2protects cells from STS-induced apoptosis, whereas apoptosis induced by Etoposide treatments (a DNA topoisomerase II inhibitor inducing apoptosis" . _:b504332899 . _:b504332921 . _:b504332900 . _:b18369115 . _:b504332901 . . _:b504332902 . _:b504332920 . _:b504332903 . _:b504332904 . _:b18369175 . _:b504332905 . _:b504332906 . _:b504332923 . _:b504332907 . _:b504332908 . _:b18369068 "The transcriptional activity of TEAD proteins requires their interaction with transcriptional co-activators [>>7<<], [8], [9]." . _:b504332909 . _:b504332910 . . _:b504332911 . _:b504332922 . _:b18369124 . _:b18369173 . _:b504332933 . _:b18369127 . _:b504332906 . _:b504332932 . _:b18369116 . _:b504332935 . _:b504332893 . _:b18369105 . _:b504332894 . _:b504332895 . _:b504332934 . . _:b18369136 "Indeed, previous studies in cultured cells showed that the activity of YAP AD is as potent as that of VP16 AD [>>52<<] and that a transcriptionally active form of TEAD1 recapitulates TEAD phenotypes when overexpressed [15], [40]." . _:b18369162 . _:b504332929 . _:b18369077 "Both Yki and YAP, are the effectors of the Hippo tumour suppressor pathway that restricts organ growth by keeping in check cell proliferation and promoting apoptosis in Drosophila and in mammals [17], [>>18<<]. The regulation of Yki/YAP activity is achieved through direct phosphorylation by the Warts/Large Tumour Suppressor (LATS) kinases that are activated by the upstream components of the Hippo pathway and subsequently induce Yki/YAP" . . . _:b504332928 . _:b504332919 . _:b504332931 . _:b18369159 "TEAD1 is constitutively expressed in cardiac and skeletal muscles in pigs, mice and humans [56], [57] where it regulates the expression of many skeletal muscle-specific genes that contain the M-CAT motif (TEAD1 protein binding site) [>>58<<], [59]. Its disruption in mice leads to heart defects and embryonic lethality between embryonic days 11 and 12 [60] while its overexpression in the mouse heart can induce age-dependent dysfunction [61]." . . _:b504332930 . _:b18369145 "This mutation causes the human genetic disease known as Sveinsson's chorioretinal atrophy [>>38<<], and strongly reduces both YAP/TEAD1 interaction and activity [55]." . _:b504332941 . . _:b18369109 _:b18369152 . _:b18369143 "Besides its partnership with TEAD1, YAP also interacts with and increases the activity of p73, a member of the p53 family that positively regulates pro-apoptotic genes [>>26<<], [54]. Thus, it is conceivable that the YAP/TEAD1 interaction competes with the YAP/p73 interaction resulting in a decrease in p73 pro-apoptotic activity." . . _:b504332940 . _:b18369109 _:b18369153 . . _:b18369109 _:b18369154 . _:b18369109 _:b18369155 . _:b504332943 . _:b18369167 "The anti-apoptotic features of Livin are well known [45], [>>65<<], [66]. Our Livin RNAi experiments not only confirm these previously published data, but also demonstrate that Livin up-regulation is required for TEAD1-induced apoptotic protection." . _:b18369102 "materials and methods" . _:b18369093 "a highly conserved family of intracellular proteins, the Inhibitor of Apoptosis Proteins (IAP) that suppress apoptosis induced by a variety of stimuli by binding specific intracellular proteases, primarily caspases 3, 7 and 9 [28], [>>29<<], [30]. In humans, eight family members have been identified (NAIP, c-IAP1, c-IAP2, XIAP, Survivin, Apollon, Livin and ILP2) [31], and only two in Drosophila (Diap1/2) [32]." . _:b504332942 . _:b504332960 . _:b504332961 . _:b504332962 . _:b504332937 . _:b18369164 . _:b18369170 "Indeed, TEAD1 and MEF2 physically interact through the TEA and MAD box domains but additional sequences in the activation domains of both proteins are required for in vivo association [>>59<<]. Our results (Fig. 4C,D) and published data [62], [63] suggest that, in HeLa cells, YAP overexpression or the Hippo signaling pathway does not play a role in apoptosis regulation. However, we observed that YAP up-regulation can modulate" . . _:b504332936 . _:b18369133 . _:b18369166 "The anti-apoptotic features of Livin are well known [>>45<<], [65], [66]. Our Livin RNAi experiments not only confirm these previously published data, but also demonstrate that Livin up-regulation is required for TEAD1-induced apoptotic protection." . _:b504332939 . _:b504332938 . _:b504332944 . _:b504332945 . _:b504332946 . _:b504332947 . _:b504332949 . _:b504332895 "3"^^ . _:b504332948 . _:b504332895 . _:b504332949 . _:b18369165 "However, the effect of a gain of function for wild-type TEAD proteins had not yet been explored either in these models or in cancer cell systems, since only a transcriptionally active form of TEAD was investigated in previous studies [>>40<<]. Here we show that overexpression or loss of function for the wild-type TEAD1 protein is sufficient to protect HeLa cells from induced apoptosis and can induce a significant up-regulation of both isoforms of the Livin protein that belongs" . _:b504332950 . _:b504332893 "6"^^ . _:b504332951 . _:b504332948 . _:b504332952 . _:b504332894 "4"^^ . _:b504332953 . _:b504332954 . _:b504332955 . _:b504332951 . _:b504332956 . _:b18369123 "The DNA binding domain of TEAD proteins (TEA domain) is highly conserved [9], whereas the C-terminal domain that interacts with cofactors is more variable, and can bind different TEAD-specific interactors [43], [>>44<<]. The observed specificity for Livin up-regulation favors a model in which TEAD1 likely interacts with a specific cofactor involved in Livin regulation. In vivo, the two known spliced isoforms of Livin mRNA present different antiapoptotic" . _:b18369115 "Transfected cells were treated with Staurosporine (STS) a large spectrum inhibitor of protein kinases, that triggers the release of cytochrome c [>>41<<]. Apoptotic cells were scored by two complementary detection methods: 1) the nuclei morphology, as visualized by Hoechst staining, highlighting chromatin condensation and/or fragmentation; 2) the immunodetection of activated caspase-3" . _:b504332957 . _:b18369069 "The transcriptional activity of TEAD proteins requires their interaction with transcriptional co-activators [7], [>>8<<], [9]. In Drosophila, recent studies have demonstrated that Sd interacts with Yorkie (Yki) [10], [11], [12]. Yki is the Drosophila ortholog of mammalian YAP (Yes-Associated Protein) which in vitro and in vivo is a well characterized" . _:b504332958 . _:b504332943 . _:b504332959 . _:b504332950 . _:b504332928 . _:b504332929 . _:b504332960 . _:b504332930 . _:b504332931 . _:b504332945 . _:b504332932 . . . _:b504332933 . _:b504332942 . _:b504332934 . _:b504332935 . _:b504332944 . _:b504332936 . _:b504332937 . _:b504332938 . _:b504332939 . _:b504332947 . _:b504332901 "3"^^ . _:b504332940 . _:b504332941 . _:b504332942 . _:b504332943 . _:b504332946 . _:b504332900 "3"^^ . _:b18369157 "TEAD1 is constitutively expressed in cardiac and skeletal muscles in pigs, mice and humans [>>56<<], [57] where it regulates the expression of many skeletal muscle-specific genes that contain the M-CAT motif (TEAD1 protein binding site) [58], [59]." . _:b504332903 "3"^^ . _:b504332957 . _:b504332902 "3"^^ . _:b504332956 . _:b18369066 . _:b504332897 "3"^^ . _:b504332959 . _:b18369150 . _:b504332896 "3"^^ . _:b504332958 . . _:b504332899 "3"^^ . _:b18369152 . _:b504332953 . . . "10.1371%2Fjournal.pone.0045498" . _:b504332898 "3"^^ . _:b504332952 . . _:b504332909 "2"^^ . _:b504332955 . _:b18369126 . _:b504332908 "2"^^ . _:b18369102 _:b18369108 . _:b18369157 . _:b504332954 . _:b504332911 "2"^^ . _:b18369102 _:b18369104 . _:b18369102 _:b18369105 . _:b18369102 _:b18369106 . _:b504332910 "2"^^ . _:b18369102 _:b18369107 . _:b18369142 . _:b18369128 "2G and 2H), consistent with previously published data [>>46<<], [48], [49], [50]. Importantly Livin RNAi also completely abolished the resistance to STS-induced apoptosis conferred by TEAD1 overexpression (Fig." . _:b18369102 _:b18369103 . _:b504332905 "3"^^ . _:b18369137 . _:b504332904 "3"^^ . _:b504332907 "2"^^ . _:b18369112 "Nevertheless, several of these results were not obtained using wild-type TEAD proteins, which for instance do not seem to affect proliferation when overexpressed [>>15<<], [16] and for which a putative effect on apoptosis had not yet been investigated." . . _:b504332961 . . . . . _:b504332960 . _:b504332906 "3"^^ . _:b504332917 "2"^^ . . _:b504332916 "2"^^ . . _:b18369134 "As expected, the TEA domain did not affect the activity of a TEAD1 responsive reporter, while \u039455-121C could inhibit this responsive reporter, as previously published [1], [>>9<<] (Fig. S2A). Nevertheless, we did not observe any Livin induction in response to either truncated protein (Fig. 3B), indicating that a functional and entire TEAD1 protein is required for Livin induction." . _:b504332962 . . _:b504332919 "2"^^ . _:b504332918 "2"^^ . . _:b504332952 . _:b504332913 "2"^^ . . _:b18369144 "Besides its partnership with TEAD1, YAP also interacts with and increases the activity of p73, a member of the p53 family that positively regulates pro-apoptotic genes [26], [>>54<<]. Thus, it is conceivable that the YAP/TEAD1 interaction competes with the YAP/p73 interaction resulting in a decrease in p73 pro-apoptotic activity." . . _:b504332933 . _:b504332912 "2"^^ . _:b504332915 "2"^^ . . _:b504332914 "2"^^ . _:b18369109 _:b18369136 . _:b18369109 _:b18369137 . _:b18369065 . _:b18369109 _:b18369138 . . _:b18369109 _:b18369139 . _:b504332925 "2"^^ . _:b18369109 _:b18369140 . _:b18369109 _:b18369141 . . _:b18369110 "active form of Tead2, (a chimeric TEA-containing the N-terminal region of Tead2 fused to the exogenous transactivation domain of VP16) in the mouse fibroblast NIH3T3 cell line, protected cells from apoptosis induced by Taxol [>>40<<]. In vivo studies showed that loss of function of TEAD1 (using a dominant negative containing the TEA domain and the surrounding sequences of TEAD1 only, but lacking the YAP-binding domain) leads to increased cell death in mouse neuronal" . _:b18369109 _:b18369142 . _:b18369089 "instance Cyclin E is induced by Yki overexpression in Drosophila [19], but not by YAP overexpression in mammalian cells where cyclin D1 is upregulated in response to a gain of function for YAP/TEAD, in mouse neural progenitor cells [>>15<<]. Moreover, some of the functions of YAP are opposite to those of Yki." . . _:b18369109 _:b18369143 . _:b18369077 . _:b504332924 "2"^^ . _:b18369109 _:b18369144 . _:b18369109 _:b18369145 . _:b18369109 _:b18369146 . _:b504332927 "2"^^ . _:b18369109 _:b18369147 . _:b18369109 _:b18369148 . _:b18369109 _:b18369149 . _:b18369109 _:b18369150 . _:b504332926 "2"^^ . _:b18369109 _:b18369151 . _:b18369109 _:b18369120 . _:b18369109 _:b18369121 . _:b18369109 _:b18369122 . _:b504332921 "2"^^ . _:b18369109 _:b18369123 . _:b18369109 _:b18369124 . _:b18369109 _:b18369125 . _:b18369109 _:b18369126 . . _:b504332920 "2"^^ . _:b18369109 _:b18369127 . _:b18369109 _:b18369128 . _:b18369109 _:b18369129 . _:b18369109 _:b18369130 . _:b504332923 "2"^^ . _:b18369109 _:b18369131 . . _:b18369109 _:b18369132 . _:b18369109 _:b18369133 . _:b18369109 _:b18369134 . _:b504332922 "2"^^ . _:b18369109 _:b18369135 . _:b504332910 . _:b504332933 "2"^^ . _:b18369109 _:b18369110 . _:b504332932 "2"^^ . _:b18369109 _:b18369111 . _:b18369072 . _:b18369109 _:b18369112 . _:b18369098 "TEAD1 has been found either upregulated, for instance in prostatic or pancreatic cancers [33], [34], or conversely decreased in bladder or breast cancer, for example (as reported by the ONCOMINE database [35], [>>36<<], [37]). Nevertheless the functional outcome and significance of such TEAD1 modulations, as well as its bona fide target genes relevant to tumorigenesis remained elusive." . _:b18369109 _:b18369113 . _:b18369100 . _:b18369109 _:b18369114 . _:b504332935 "2"^^ . _:b18369109 _:b18369115 . _:b18369109 _:b18369116 . _:b18369109 _:b18369117 . _:b18369109 _:b18369118 . _:b504332934 "2"^^ . _:b18369109 _:b18369119 . _:b18369118 . _:b504332929 "2"^^ . _:b504332928 "2"^^ . _:b18369112 . _:b504332931 "2"^^ . _:b504332949 . _:b504332930 "2"^^ . _:b18369107 . _:b504332953 . _:b504332941 "2"^^ . _:b18369103 . _:b504332940 "2"^^ . _:b504332914 . . . _:b504332943 "2"^^ . _:b18369111 . _:b504332942 "2"^^ . _:b504332904 . _:b504332937 "2"^^ . _:b18369152 "of such highly limiting, possibly cell-specific, titratable transcriptional co-factor(s) has been deduced from transfection analyses where activation of a cognate reporter is severely reduced upon TEAD1 overexpression [1], [9], [>>56<<], [57], [58] (Fig. S2A)." . _:b504332936 "2"^^ . _:b18369131 "2G and 2H), consistent with previously published data [46], [48], [49], [>>50<<]. Importantly Livin RNAi also completely abolished the resistance to STS-induced apoptosis conferred by TEAD1 overexpression (Fig." . _:b504332939 "2"^^ . . _:b504332938 "2"^^ . _:b504332949 "2"^^ . _:b18369061 "introduction" . _:b18369123 . _:b504332948 "2"^^ . _:b18369153 "of such highly limiting, possibly cell-specific, titratable transcriptional co-factor(s) has been deduced from transfection analyses where activation of a cognate reporter is severely reduced upon TEAD1 overexpression [1], [9], [56], [>>57<<], [58] (Fig. S2A)." . _:b18369106 . . . _:b504332951 "2"^^ . _:b504332909 . _:b504332937 . _:b18369111 "that loss of function of TEAD1 (using a dominant negative containing the TEA domain and the surrounding sequences of TEAD1 only, but lacking the YAP-binding domain) leads to increased cell death in mouse neuronal progenitor cells [>>15<<]. Nevertheless, several of these results were not obtained using wild-type TEAD proteins, which for instance do not seem to affect proliferation when overexpressed [15], [16] and for which a putative effect on apoptosis had not yet been" . _:b504332950 "2"^^ . _:b18369071 "In Drosophila, recent studies have demonstrated that Sd interacts with Yorkie (Yki) [>>10<<], [11], [12]." . _:b504332945 "2"^^ . _:b504332944 "2"^^ . _:b18369096 . _:b504332947 "2"^^ . _:b504332954 . _:b18369062 "TEAD1 belongs to the family of conserved eukaryotic transcription factors (TEAD proteins), characterized by the TEA/ATTS DNA binding domain [>>1<<], [2], [3]. There are four closely related Tead genes (Tead1 to Tead4) in mammals [4], [5] and one, scalloped (sd), in Drosophila [6]." . _:b504332946 "2"^^ . _:b504332957 "2"^^ . . _:b18369096 "TEAD1 has been found either upregulated, for instance in prostatic or pancreatic cancers [>>33<<], [34], or conversely decreased in bladder or breast cancer, for example (as reported by the ONCOMINE database [35], [36], [37])." . . _:b504332956 "2"^^ . _:b18369097 "TEAD1 has been found either upregulated, for instance in prostatic or pancreatic cancers [33], [>>34<<], or conversely decreased in bladder or breast cancer, for example (as reported by the ONCOMINE database [35], [36], [37])." . _:b504332959 "2"^^ . _:b504332958 "2"^^ . _:b18369127 "[47] was blocked only by the Livin \u03B2 isoform. Since RT-Q-PCR cannot distinguish between these two splice variants, semi-quantitative PCR experiments with primers flanking the truncated region of exon 6 were performed in parallel. [>>45<<]. Our results indicate a TEAD1-dependent induction of both \u03B1 and \u03B2Livin isoforms (Fig." . _:b18369063 . _:b504332953 "2"^^ . _:b18369089 . _:b18369098 . _:b504332952 "2"^^ . _:b18369150 "The existence of such highly limiting, possibly cell-specific, titratable transcriptional co-factor(s) has been deduced from transfection analyses where activation of a cognate reporter is severely reduced upon TEAD1 overexpression [>>1<<], [9], [56], [57], [58] (Fig. S2A)." . _:b18369062 . _:b504332955 "2"^^ . _:b18369178 . _:b504332954 "2"^^ . _:b18369156 _:b18369172 . _:b18369156 _:b18369173 . _:b18369156 _:b18369174 . _:b18369156 _:b18369175 . _:b18369156 _:b18369168 . _:b18369156 _:b18369169 . _:b18369156 _:b18369170 . _:b18369156 _:b18369171 . . _:b18369156 _:b18369176 . _:b18369156 _:b18369177 . _:b18369156 _:b18369178 . _:b18369156 _:b18369179 . _:b18369156 _:b18369157 . _:b18369156 _:b18369158 . _:b18369156 _:b18369159 . _:b504332961 "2"^^ . _:b504332960 "2"^^ . _:b18369172 "4C,D) and published data [62], [>>63<<] suggest that, in HeLa cells, YAP overexpression or the Hippo signaling pathway does not play a role in apoptosis regulation." . _:b18369156 _:b18369164 . _:b18369140 "As a positive control, we quantified the mRNA levels of CTGF, a gene known to be directly activated by TEAD1 and YAP [>>13<<]. As expected, CTGF expression was induced by TEAD-VP16 in a dose-dependent manner (Fig. 4A). Conversely, TEAD-VP16 did not affect Livin mRNA expression (Fig. 4B), in accordance with results obtained with the Livin promoter. Nevertheless," . _:b18369156 _:b18369165 . _:b18369156 _:b18369166 . _:b18369156 _:b18369167 . _:b18369156 _:b18369160 . _:b18369156 _:b18369161 . _:b18369108 "Vucic) contains a functional 1.6 kb region of the promoter of Livin cloned into the pGL4.10 plasmid [>>51<<]. The \u22122570/\u22122518-TK164-luc plasmid (a gift of A. Payne) contains a 53 bp fragment from the promoter of the 3\u03B2-hydroxysteroid dehydrogenase-isomerase (3\u03B2HSD) promoter cloned upstream of a thymidine kinase minimal promoter. This 53 bp" . _:b504332900 . _:b18369156 _:b18369162 . _:b18369156 _:b18369163 . _:b504332962 "2"^^ . _:b504332915 . . . . _:b18369174 . . _:b18369156 . _:b18369138 "Indeed, previous studies in cultured cells showed that the activity of YAP AD is as potent as that of VP16 AD [52] and that a transcriptionally active form of TEAD1 recapitulates TEAD phenotypes when overexpressed [15], [>>40<<]. As expected, the TEAD-VP16 fusion protein stimulated a TEAD-responsive reporter construct in a dose-dependent manner (Fig. S2B) while the TEAD-ENRD fusion protein strongly repressed that same reporter [53] (Fig. S2A)." . _:b18369095 "In humans, eight family members have been identified (NAIP, c-IAP1, c-IAP2, XIAP, Survivin, Apollon, Livin and ILP2) [31], and only two in Drosophila (Diap1/2) [>>32<<]." . _:b504332961 . _:b18369073 . . . _:b18369074 . . _:b504332916 . _:b18369126 "cells as a model, Ashhab et al. (2001) showed that Livin \u03B1but not \u03B2protects cells from STS-induced apoptosis, whereas apoptosis induced by Etoposide treatments (a DNA topoisomerase II inhibitor inducing apoptosis through DNA damage) [>>47<<] was blocked only by the Livin \u03B2 isoform. Since RT-Q-PCR cannot distinguish between these two splice variants, semi-quantitative PCR experiments with primers flanking the truncated region of exon 6 were performed in parallel." . _:b18369082 . . . _:b18369088 "For instance Cyclin E is induced by Yki overexpression in Drosophila [>>19<<], but not by YAP overexpression in mammalian cells where cyclin D1 is upregulated in response to a gain of function for YAP/TEAD, in mouse neural progenitor cells [15]." . . _:b18369080 . _:b18369083 "This is achieved in Drosophila through the transcriptional induction of target genes including Cyclin E, dE2F1 [>>10<<], the bantam microRNA [22], [23], dmyc [24], [25] and the Drosophila inhibitor of apoptosis protein 1 (Diap1) [17]." . . _:b504332931 . . . _:b18369145 . . _:b18369061 . . _:b18369062 . _:b18369172 . _:b18369063 . . _:b18369109 "results" . . _:b18369073 "Yki is the Drosophila ortholog of mammalian YAP (Yes-Associated Protein) which in vitro and in vivo is a well characterized cofactor of the mammalian TEAD proteins [13], [>>14<<], [15], [16]." . _:b18369176 "In agreement with our results, it has been shown that in cancers where TEAD1 is overexpressed, such as in prostate cancers [33], Livin is also up-regulated [>>64<<], [65]. However, interestingly, a modest but significant, increase in Livin is also observed in other types of cancers where TEAD1 is down-regulated, such as breast, renal or bladder cancers (www.oncomine.org). The data presented in our" . _:b18369102 . _:b504332955 . _:b18369075 . _:b18369066 "There are four closely related Tead genes (Tead1 to Tead4) in mammals [4], [>>5<<] and one, scalloped (sd), in Drosophila [6]." . _:b18369086 . _:b18369068 . _:b18369100 "Interestingly, we show that overexpression of a mutated form of TEAD1 (Y421H) implicated in Sveinsson's chorioretinal atrophy that strongly reduces interaction of the mutant with YAP as well as its activation [>>38<<], [39] can induce Livin expression and protect cells from induced apoptosis, suggesting that YAP is not the cofactor involved in Livin repression." . _:b18369069 . . _:b18369109 . _:b18369070 . _:b18369071 . . _:b504332921 . . _:b18369087 "This is achieved in Drosophila through the transcriptional induction of target genes including Cyclin E, dE2F1 [10], the bantam microRNA [22], [23], dmyc [24], [25] and the Drosophila inhibitor of apoptosis protein 1 (Diap1) [>>17<<]. However, although the framework of the Hippo signaling cascade is conserved between Drosophila and mammals there is still significant ambiguity as to how the pathway converges onto transcriptional regulators and elicits coherent" . . _:b18369064 . _:b18369076 . . . _:b18369065 . _:b18369061 . . _:b18369066 . _:b504332899 . _:b18369067 . _:b18369067 . _:b18369076 . _:b18369165 . _:b18369163 "Moreover, recent data support an anti-apoptotic role for TEAD proteins during mouse development, based on Tead1 and/or Tead2 loss of function experiments [>>15<<], [16]. However, the effect of a gain of function for wild-type TEAD proteins had not yet been explored either in these models or in cancer cell systems, since only a transcriptionally active form of TEAD was investigated in previous" . . _:b18369094 "conserved family of intracellular proteins, the Inhibitor of Apoptosis Proteins (IAP) that suppress apoptosis induced by a variety of stimuli by binding specific intracellular proteases, primarily caspases 3, 7 and 9 [28], [29], [>>30<<]. In humans, eight family members have been identified (NAIP, c-IAP1, c-IAP2, XIAP, Survivin, Apollon, Livin and ILP2) [31], and only two in Drosophila (Diap1/2) [32]." . . _:b18369077 . . _:b18369078 . . _:b18369141 "To identify the putative cofactor involved in this activation, we further investigated the transcriptional co-activator YAP, one of the best-described cofactors for TEAD1 [>>13<<], [15]. YAP activity is regulated through its shuttling between the nucleus and the cytoplasm, under the control of the Hippo pathway." . . _:b18369079 . . _:b18369072 . _:b18369073 . _:b18369087 . _:b18369074 . _:b18369106 "Davidson) [>>1<<], TEAD1-Y421H, and pEGFP (GFP) (Clontech) cDNAs were subcloned into the pXJ40 vector; the YAP1, MST2 and LAST1 cDNAs (gifts from X." . _:b18369075 . _:b18369177 . _:b18369084 . . _:b18369085 . _:b18369097 . _:b18369086 . . _:b18369087 . _:b18369138 . _:b18369067 "There are four closely related Tead genes (Tead1 to Tead4) in mammals [4], [5] and one, scalloped (sd), in Drosophila [>>6<<]. The transcriptional activity of TEAD proteins requires their interaction with transcriptional co-activators [7], [8], [9]. In Drosophila, recent studies have demonstrated that Sd interacts with Yorkie (Yki) [10], [11], [12]. Yki is the" . _:b18369080 . _:b18369155 . _:b18369081 . _:b18369120 "novel, since a similar effect had previously been observed with a TEAD chimera fused with an exogenous transcriptional activation domain (Tead2-VP16), in another cell type (mouse fibroblasts) and using Taxol as inducer of apoptosis [>>40<<]. Taken together, our observations indicate that the alteration of wild-type TEAD1 expression levels is sufficient to promote a cytoprotective effect against pro-apoptotic stimuli." . _:b18369091 "YAP, as a cofactor for p73 (a member of the p53 family of transcription factors) can promote apoptosis after DNA damage [26], [>>27<<], whereas Yki is clearly a suppressor of cell death in the fly eye." . _:b18369082 . _:b18369083 . _:b18369125 . _:b18369113 "Nevertheless, several of these results were not obtained using wild-type TEAD proteins, which for instance do not seem to affect proliferation when overexpressed [15], [>>16<<] and for which a putative effect on apoptosis had not yet been investigated." . _:b18369092 . _:b18369093 . . . _:b18369094 . . . _:b18369078 "is achieved through direct phosphorylation by the Warts/Large Tumour Suppressor (LATS) kinases that are activated by the upstream components of the Hippo pathway and subsequently induce Yki/YAP cytoplasmic retention and inactivation [>>19<<], [20], [21]. Conversely,Yki overexpression promotes organ growth by stimulating cell proliferation and preventing apoptosis [19], [21]." . . _:b18369095 . _:b18369088 . . _:b18369089 . _:b18369090 . _:b18369135 . _:b18369091 . _:b18369170 . _:b504332922 . . _:b18369100 . _:b18369101 . _:b504332902 . . _:b18369167 . _:b18369102 . _:b18369090 . _:b18369103 . _:b18369096 . _:b18369128 . _:b504332930 . _:b18369097 . _:b18369120 . _:b18369134 . _:b18369166 . _:b18369098 . _:b18369099 . _:b504332950 . _:b18369108 . _:b504332928 . _:b18369109 . _:b18369110 . _:b18369068 . _:b18369111 . . _:b504332958 . _:b18369104 . _:b18369179 . _:b504332917 . _:b504332896 . _:b18369105 . _:b18369069 . . . _:b18369106 . . _:b18369154 . _:b18369110 . _:b18369107 . _:b18369116 . _:b18369117 . _:b18369107 "Thiebaud) [>>68<<]. HeLa, BUA and MCF7 cells were transfected with these plasmids (individually or in combination), together with GFP, using the Effectene Transfection Reagent (Qiagen), according to the manufacturer's instructions." . _:b18369168 . _:b18369122 . . _:b18369159 . _:b18369118 . _:b18369121 . _:b18369075 "Yki is the Drosophila ortholog of mammalian YAP (Yes-Associated Protein) which in vitro and in vivo is a well characterized cofactor of the mammalian TEAD proteins [13], [14], [15], [>>16<<]. Both Yki and YAP, are the effectors of the Hippo tumour suppressor pathway that restricts organ growth by keeping in check cell proliferation and promoting apoptosis in Drosophila and in mammals [17], [18]. The regulation of Yki/YAP" . _:b18369119 . _:b18369112 . . _:b18369139 . _:b18369151 . . . _:b18369072 "Yki is the Drosophila ortholog of mammalian YAP (Yes-Associated Protein) which in vitro and in vivo is a well characterized cofactor of the mammalian TEAD proteins [>>13<<], [14], [15], [16]." . _:b18369160 . _:b18369113 . _:b18369101 . _:b18369114 . . _:b18369149 . . _:b18369146 . _:b18369115 . _:b18369164 "Moreover, recent data support an anti-apoptotic role for TEAD proteins during mouse development, based on Tead1 and/or Tead2 loss of function experiments [15], [>>16<<]. However, the effect of a gain of function for wild-type TEAD proteins had not yet been explored either in these models or in cancer cell systems, since only a transcriptionally active form of TEAD was investigated in previous studies" . _:b504332945 . . _:b18369124 . _:b18369125 . _:b18369091 . _:b18369147 "Yet, even if it is well established that TEAD1 must interact with cofactors to activate transcription [>>7<<], [8], TEAD1 overexpression in cultured cells may inhibit their transcriptional functions by \u201Ctitrating\u201D co-activator proteins [9]." . _:b18369126 . _:b18369132 . _:b18369127 . _:b18369120 . _:b504332936 . . _:b18369121 . _:b18369122 . _:b504332897 . _:b18369123 . _:b18369153 . _:b18369085 "This is achieved in Drosophila through the transcriptional induction of target genes including Cyclin E, dE2F1 [10], the bantam microRNA [22], [>>23<<], dmyc [24], [25] and the Drosophila inhibitor of apoptosis protein 1 (Diap1) [17]." . _:b18369174 "Although its transcriptional regulation is poorly understood, TEAD1 is up-regulated in several types of cancers such as prostatic or pancreatic cancers [33], [>>34<<], reaching up to 300 fold induction in Kaposi's sarcoma, (Gene Expression Atlas :" . _:b18369132 . . _:b18369070 "The transcriptional activity of TEAD proteins requires their interaction with transcriptional co-activators [7], [8], [>>9<<]. In Drosophila, recent studies have demonstrated that Sd interacts with Yorkie (Yki) [10], [11], [12]. Yki is the Drosophila ortholog of mammalian YAP (Yes-Associated Protein) which in vitro and in vivo is a well characterized cofactor of" . _:b18369133 . _:b18369137 "Indeed, previous studies in cultured cells showed that the activity of YAP AD is as potent as that of VP16 AD [52] and that a transcriptionally active form of TEAD1 recapitulates TEAD phenotypes when overexpressed [>>15<<], [40]. As expected, the TEAD-VP16 fusion protein stimulated a TEAD-responsive reporter construct in a dose-dependent manner (Fig. S2B) while the TEAD-ENRD fusion protein strongly repressed that same reporter [53] (Fig. S2A)." . . _:b18369134 . . . _:b18369135 . _:b18369148 "Yet, even if it is well established that TEAD1 must interact with cofactors to activate transcription [7], [>>8<<], TEAD1 overexpression in cultured cells may inhibit their transcriptional functions by \u201Ctitrating\u201D co-activator proteins [9]." . _:b18369084 "This is achieved in Drosophila through the transcriptional induction of target genes including Cyclin E, dE2F1 [10], the bantam microRNA [>>22<<], [23], dmyc [24], [25] and the Drosophila inhibitor of apoptosis protein 1 (Diap1) [17]." . _:b18369128 . _:b504332894 . _:b18369129 . _:b18369125 "In vivo, the two known spliced isoforms of Livin mRNA present different antiapoptotic properties [45], [>>46<<]. Using Jurkat cells as a model, Ashhab et al. (2001) showed that Livin \u03B1but not \u03B2protects cells from STS-induced apoptosis, whereas apoptosis induced by Etoposide treatments (a DNA topoisomerase II inhibitor inducing apoptosis through DNA" . _:b18369117 "S1C), reducing the risk of a possible functional redundancy among TEAD paralogs that could compensate for the absence or reduction of TEAD1 [>>16<<]. Specific mRNA knockdown by two independent synthetic siRNAs (TEAD1-5 and TEAD1-8) reduced TEAD1 to undetectable levels in Western blots (Fig. 1B). Treatment of cells with either of the TEAD1 siRNAs significantly protected HeLa cells from" . _:b18369130 . _:b18369158 . . _:b18369131 . _:b504332934 . _:b18369080 "through direct phosphorylation by the Warts/Large Tumour Suppressor (LATS) kinases that are activated by the upstream components of the Hippo pathway and subsequently induce Yki/YAP cytoplasmic retention and inactivation [19], [20], [>>21<<]. Conversely,Yki overexpression promotes organ growth by stimulating cell proliferation and preventing apoptosis [19], [21]." . . _:b18369140 . _:b18369071 . _:b18369141 . _:b18369158 "TEAD1 is constitutively expressed in cardiac and skeletal muscles in pigs, mice and humans [56], [>>57<<] where it regulates the expression of many skeletal muscle-specific genes that contain the M-CAT motif (TEAD1 protein binding site) [58], [59]." . _:b18369142 . _:b18369092 . _:b18369143 . _:b18369090 "YAP, as a cofactor for p73 (a member of the p53 family of transcription factors) can promote apoptosis after DNA damage [>>26<<], [27], whereas Yki is clearly a suppressor of cell death in the fly eye." . _:b18369136 . _:b18369137 . _:b18369084 . . _:b18369138 . _:b504332924 . _:b18369139 . . _:b18369148 . "PMC0" . _:b18369149 . _:b18369150 . . _:b504332932 . . _:b18369151 . _:b18369079 "through direct phosphorylation by the Warts/Large Tumour Suppressor (LATS) kinases that are activated by the upstream components of the Hippo pathway and subsequently induce Yki/YAP cytoplasmic retention and inactivation [19], [>>20<<], [21]. Conversely,Yki overexpression promotes organ growth by stimulating cell proliferation and preventing apoptosis [19], [21]." . _:b504332920 . _:b18369144 . . _:b18369083 . _:b18369171 . _:b18369133 "As expected, the TEA domain did not affect the activity of a TEAD1 responsive reporter, while \u039455-121C could inhibit this responsive reporter, as previously published [>>1<<], [9] (Fig." . _:b18369145 . _:b18369179 "Although Vgll1 and YAP are not homologous, a recent study has shown that Vgll1 interacts with TEAD by sharing two out of the three interfaces required for the interaction with YAP [>>66<<]. In addition, a competition was observed between Vgll1 and YAP for their binding to TEAD. However, the authors have shown that some of genes regulated by YAP/TEAD, such as IGFBP-3, SERPINE1 or FGF1, are not up-regulated by Vgll1." . . _:b18369146 . . _:b18369147 . . _:b18369143 . _:b18369063 "TEAD1 belongs to the family of conserved eukaryotic transcription factors (TEAD proteins), characterized by the TEA/ATTS DNA binding domain [1], [>>2<<], [3]. There are four closely related Tead genes (Tead1 to Tead4) in mammals [4], [5] and one, scalloped (sd), in Drosophila [6]." . _:b18369156 . _:b18369157 . . _:b18369158 . _:b18369085 . _:b18369159 . _:b504332947 . _:b18369173 "Although its transcriptional regulation is poorly understood, TEAD1 is up-regulated in several types of cancers such as prostatic or pancreatic cancers [>>33<<], [34], reaching up to 300 fold induction in Kaposi's sarcoma, (Gene Expression Atlas :" . _:b18369079 . _:b504332926 . _:b18369161 . . _:b18369152 . _:b18369148 . . _:b18369099 "TEAD1 has been found either upregulated, for instance in prostatic or pancreatic cancers [33], [34], or conversely decreased in bladder or breast cancer, for example (as reported by the ONCOMINE database [35], [36], [>>37<<]). Nevertheless the functional outcome and significance of such TEAD1 modulations, as well as its bona fide target genes relevant to tumorigenesis remained elusive." . _:b18369153 . _:b18369149 "Yet, even if it is well established that TEAD1 must interact with cofactors to activate transcription [7], [8], TEAD1 overexpression in cultured cells may inhibit their transcriptional functions by \u201Ctitrating\u201D co-activator proteins [>>9<<]. The existence of such highly limiting, possibly cell-specific, titratable transcriptional co-factor(s) has been deduced from transfection analyses where activation of a cognate reporter is severely reduced upon TEAD1 overexpression [1]," . _:b504332962 . _:b18369154 . . _:b18369155 . _:b18369082 "Conversely,Yki overexpression promotes organ growth by stimulating cell proliferation and preventing apoptosis [19], [>>21<<]. This is achieved in Drosophila through the transcriptional induction of target genes including Cyclin E, dE2F1 [10], the bantam microRNA [22], [23], dmyc [24], [25] and the Drosophila inhibitor of apoptosis protein 1 (Diap1) [17]." . _:b18369164 . _:b18369155 "As positive control, the mRNA of the TEAD1-YAP target gene CTGF [>>13<<] was quantified." . _:b18369147 . _:b18369165 . . _:b504332938 . . _:b18369108 . _:b18369166 . _:b18369135 "TEAD1-induced apoptotic resistance and Livin induction are mediated through the transcriptional activity of TEAD1, a constitutively active form and a repressive form of TEAD1 were constructed by replacing its YAP binding domain [>>14<<] by the activation domain (AD) of VP16 (TEAD-VP16) or the repressor domain (RD) of Engrailed (TEAD-ENRD) (Fig." . _:b18369122 "The DNA binding domain of TEAD proteins (TEA domain) is highly conserved [>>9<<], whereas the C-terminal domain that interacts with cofactors is more variable, and can bind different TEAD-specific interactors [43], [44]." . _:b18369065 "There are four closely related Tead genes (Tead1 to Tead4) in mammals [>>4<<], [5] and one, scalloped (sd), in Drosophila [6]." . _:b18369167 . _:b18369160 "is constitutively expressed in cardiac and skeletal muscles in pigs, mice and humans [56], [57] where it regulates the expression of many skeletal muscle-specific genes that contain the M-CAT motif (TEAD1 protein binding site) [58], [>>59<<]. Its disruption in mice leads to heart defects and embryonic lethality between embryonic days 11 and 12 [60] while its overexpression in the mouse heart can induce age-dependent dysfunction [61]." . . _:b18369176 . _:b18369161 "Its disruption in mice leads to heart defects and embryonic lethality between embryonic days 11 and 12 [>>60<<] while its overexpression in the mouse heart can induce age-dependent dysfunction [61]." . _:b18369160 . _:b18369161 . _:b18369070 . _:b18369076 "Both Yki and YAP, are the effectors of the Hippo tumour suppressor pathway that restricts organ growth by keeping in check cell proliferation and promoting apoptosis in Drosophila and in mammals [>>17<<], [18]. The regulation of Yki/YAP activity is achieved through direct phosphorylation by the Warts/Large Tumour Suppressor (LATS) kinases that are activated by the upstream components of the Hippo pathway and subsequently induce Yki/YAP" . _:b18369113 . _:b18369162 . _:b18369114 . _:b18369163 . _:b18369142 "To identify the putative cofactor involved in this activation, we further investigated the transcriptional co-activator YAP, one of the best-described cofactors for TEAD1 [13], [>>15<<]. YAP activity is regulated through its shuttling between the nucleus and the cytoplasm, under the control of the Hippo pathway." . _:b18369172 . _:b504332907 . _:b18369064 . _:b18369173 . _:b504332929 . _:b18369117 . . _:b18369174 . _:b18369144 . . _:b18369175 . _:b18369119 . _:b18369168 . _:b504332903 . _:b504332956 . _:b18369156 "discussion" . _:b504332911 . _:b18369169 . _:b504332935 . _:b18369170 . _:b504332940 . _:b18369171 . _:b18369129 "2G and 2H), consistent with previously published data [46], [>>48<<], [49], [50]. Importantly Livin RNAi also completely abolished the resistance to STS-induced apoptosis conferred by TEAD1 overexpression (Fig." . _:b504332908 . _:b18369088 . . _:b504332913 . . _:b18369176 . _:b504332941 . _:b18369177 . _:b18369178 .