FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2004102023> ?p ?o ?g. }

• W2004102023 endingPage "410" @default.
• W2004102023 startingPage "398" @default.
• W2004102023 abstract "Outside cells of the preimplantation mouse embryo form the trophectoderm (TE), a process requiring the transcription factor Tead4. Here, we show that transcriptionally active Tead4 can induce Cdx2 and other trophoblast genes in parallel in embryonic stem cells. In embryos, the Tead4 coactivator protein Yap localizes to nuclei of outside cells, and modulation of Tead4 or Yap activity leads to changes in Cdx2 expression. In inside cells, Yap is phosphorylated and cytoplasmic, and this involves the Hippo signaling pathway component Lats. We propose that active Tead4 promotes TE development in outside cells, whereas Tead4 activity is suppressed in inside cells by cell contact- and Lats-mediated inhibition of nuclear Yap localization. Thus, differential signaling between inside and outside cell populations leads to changes in cell fate specification during TE formation. Outside cells of the preimplantation mouse embryo form the trophectoderm (TE), a process requiring the transcription factor Tead4. Here, we show that transcriptionally active Tead4 can induce Cdx2 and other trophoblast genes in parallel in embryonic stem cells. In embryos, the Tead4 coactivator protein Yap localizes to nuclei of outside cells, and modulation of Tead4 or Yap activity leads to changes in Cdx2 expression. In inside cells, Yap is phosphorylated and cytoplasmic, and this involves the Hippo signaling pathway component Lats. We propose that active Tead4 promotes TE development in outside cells, whereas Tead4 activity is suppressed in inside cells by cell contact- and Lats-mediated inhibition of nuclear Yap localization. Thus, differential signaling between inside and outside cell populations leads to changes in cell fate specification during TE formation. During mouse development, the first lineage specified is the trophoblast/placenta lineage, set aside during blastocyst formation. In the blastocyst, the trophoblast, or trophectoderm (TE), surrounds the inner cell mass (ICM), which will give rise to the fetus and other extraembryonic tissues. The homeodomain transcription factor Cdx2 is expressed in the TE at the blastocyst stage. Cdx2 is required for TE development and is sufficient to promote trophoblast fate in ICM-derived embryonic stem (ES) cells, including suppression of ICM and induction of TE genes (Niwa et al., 2005Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation.Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (838) Google Scholar, Strumpf et al., 2005Strumpf D. Mao C.A. Yamanaka Y. Ralston A. Chawengsaksophak K. Beck F. Rossant J. Cdx2 is required for correct cell fate specification and differentiation of trophectoderm in the mouse blastocyst.Development. 2005; 132: 2093-2102Crossref PubMed Scopus (775) Google Scholar). Conversely, ICM fates are regulated by a distinct set of transcription factors, including the POU family transcription factor Oct3/4 (encoded by Pou5f1). Prior to blastocyst formation, Cdx2 and Oct3/4 are initially coexpressed throughout the embryo (Dietrich and Hiiragi, 2007Dietrich J.E. Hiiragi T. Stochastic patterning in the mouse pre-implantation embryo.Development. 2007; 134: 4219-4231Crossref PubMed Scopus (375) Google Scholar, Palmieri et al., 1994Palmieri S.L. Peter W. Hess H. Scholer H.R. Oct-4 transcription factor is differentially expressed in the mouse embryo during establishment of the first two extraembryonic cell lineages involved in implantation.Dev. Biol. 1994; 166: 259-267Crossref PubMed Scopus (514) Google Scholar, Ralston and Rossant, 2008Ralston A. Rossant J. Cdx2 acts downstream of cell polarization to cell-autonomously promote trophectoderm fate in the early mouse embryo.Dev. Biol. 2008; 313: 614-629Crossref PubMed Scopus (251) Google Scholar). Mutual antagonism between these two factors may contribute to the eventual segregation of their expression domains (Niwa et al., 2005Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation.Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (838) Google Scholar), with Cdx2 in outside cells of the TE and Oct3/4 in inside cells of the ICM. However, molecular mechanisms that initially interpret inside/outside positional information within the embryo to establish this pattern are not known. We and others recently showed that the TEAD/TEF family transcription factor Tead4 is essential for TE development and Cdx2 expression prior to the blastocyst stage (Nishioka et al., 2008Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos.Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (318) Google Scholar, Yagi et al., 2007Yagi R. Kohn M.J. Karavanova I. Kaneko K.J. Vullhorst D. Depamphilis M.L. Buonanno A. Transcription factor TEAD4 specifies the trophectoderm lineage at the beginning of mammalian development.Development. 2007; 134: 3827-3836Crossref PubMed Scopus (339) Google Scholar). This provided the first clue about molecular mechanisms acting upstream of the TE/ICM lineage distinction. However, whether Tead4 acted permissively or instructively in this process was unclear, since Tead4 itself was not restricted to outside cells (Nishioka et al., 2008Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos.Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (318) Google Scholar). Here, we sought to identify cofactors and signaling components that could impart positional information to spatially regulate Tead4 activity in the embryo. Many lines of evidence have suggested that TEAD-mediated transcription is regulated by the Ser/Thr kinase Hippo in Drosophila, or Stk3 (Mst) in mice. In Drosophila, Hippo inhibits the Yorkie (Yki) coactivator and suppresses cell proliferation (Huang et al., 2005Huang J. Wu S. Barrera J. Matthews K. Pan D. The Hippo signaling pathway coordinately regulates cell proliferation and apoptosis by inactivating Yorkie, the Drosophila homolog of YAP.Cell. 2005; 122: 421-434Abstract Full Text Full Text PDF PubMed Scopus (1179) Google Scholar). These activities are mediated by a Tead protein, Scalloped (Goulev et al., 2008Goulev Y. Fauny J.D. Gonzalez-Marti B. Flagiello D. Silber J. Zider A. SCALLOPED interacts with YORKIE, the nuclear effector of the hippo tumor-suppressor pathway in Drosophila.Curr. Biol. 2008; 18: 435-441Abstract Full Text Full Text PDF PubMed Scopus (282) Google Scholar, Wu et al., 2008Wu S. Liu Y. Zheng Y. Dong J. Pan D. The TEAD/TEF family protein Scalloped mediates transcriptional output of the Hippo growth-regulatory pathway.Dev. Cell. 2008; 14: 388-398Abstract Full Text Full Text PDF PubMed Scopus (452) Google Scholar, Zhang et al., 2008bZhang L. Ren F. Zhang Q. Chen Y. Wang B. Jiang J. The TEAD/TEF family of transcription factor Scalloped mediates Hippo signaling in organ size control.Dev. Cell. 2008; 14: 377-387Abstract Full Text Full Text PDF PubMed Scopus (438) Google Scholar). In mammals, Hippo signaling comprises a growth-regulating pathway, which controls cell contact-mediated inhibition of proliferation (see reviews Pan, 2007Pan D. Hippo signaling in organ size control.Genes Dev. 2007; 21: 886-897Crossref PubMed Scopus (476) Google Scholar, Reddy and Irvine, 2008Reddy B.V. Irvine K.D. The Fat and Warts signaling pathways: new insights into their regulation, mechanism and conservation.Development. 2008; 135: 2827-2838Crossref PubMed Scopus (162) Google Scholar, Saucedo and Edgar, 2007Saucedo L.J. Edgar B.A. Filling out the Hippo pathway.Nat. Rev. Mol. Cell Biol. 2007; 8: 613-621Crossref PubMed Scopus (261) Google Scholar). In this context, cell-cell contact regulates nuclear accumulation of a Yki homolog, Yes-associated protein 1 (Yap1, Yap hereafter), through Hippo signaling and controls cell proliferation by regulating transcriptional activity of Tead proteins (Ota and Sasaki, 2008Ota M. Sasaki H. Mammalian Tead proteins regulate cell proliferation and contact inhibition as a transcriptional mediator of Hippo signaling.Development. 2008; 135: 4059-4069Crossref PubMed Scopus (274) Google Scholar, Zhao et al., 2007Zhao B. Wei X. Li W. Udan R.S. Yang Q. Kim J. Xie J. Ikenoue T. Yu J. Li L. et al.Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control.Genes Dev. 2007; 21: 2747-2761Crossref PubMed Scopus (1767) Google Scholar, Zhao et al., 2008Zhao B. Ye X. Yu J. Li L. Li W. Li S. Yu J. Lin J.D. Wang C.Y. Chinnaiyan A.M. et al.TEAD mediates YAP-dependent gene induction and growth control.Genes Dev. 2008; 22: 1962-1971Crossref PubMed Scopus (1408) Google Scholar). In the mouse embryo, Tead1−/−; Tead2−/− mutants die soon after implantation due to reduced cell proliferation and increased apoptosis (Sawada et al., 2008Sawada A. Kiyonari H. Ukita K. Nishioka N. Imuta Y. Sasaki H. Redundant roles of Tead1 and Tead2 in notochord development and the regulation of cell proliferation and survival.Mol. Cell. Biol. 2008; 28: 3177-3189Crossref PubMed Scopus (122) Google Scholar). Tead1/2 interact genetically with Yap (Sawada et al., 2008Sawada A. Kiyonari H. Ukita K. Nishioka N. Imuta Y. Sasaki H. Redundant roles of Tead1 and Tead2 in notochord development and the regulation of cell proliferation and survival.Mol. Cell. Biol. 2008; 28: 3177-3189Crossref PubMed Scopus (122) Google Scholar), suggesting that the roles of these genes in Hippo signaling are conserved in mice. We examined the role of Tead4 in TE development using both ES cells and preimplantation embryos. We show that active Tead4 can promote multiple trophoblast genes in parallel, including Cdx2. We next show that, in the embryo, the Tead coactivator Yap localizes to nuclei only in outside cells and is excluded from inside cell nuclei by the Hippo signaling pathway component Lats. These observations suggest that Tead4/Yap interpret positional information along the inside/outside axis of the embryo to restrict expression of Cdx2 and TE fates to outside cells. Although Tead4 is required for establishment of the TE lineage in the mouse embryo (Nishioka et al., 2008Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos.Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (318) Google Scholar, Yagi et al., 2007Yagi R. Kohn M.J. Karavanova I. Kaneko K.J. Vullhorst D. Depamphilis M.L. Buonanno A. Transcription factor TEAD4 specifies the trophectoderm lineage at the beginning of mammalian development.Development. 2007; 134: 3827-3836Crossref PubMed Scopus (339) Google Scholar), the sufficiency of Tead4 to promote TE fate has not been reported. We examined the ability of Tead4 to promote trophoblast differentiation in ES cells. For comparison, we used the ES cell line 5ECER4 (Niwa et al., 2005Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation.Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (838) Google Scholar), which stably expresses a tamoxifen (Tx)-inducible fusion between Cdx2 and a modified ligand-binding domain of the estrogen receptor (ER) (Cdx2ER). Consistent with previous analyses (Niwa et al., 2005Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation.Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (838) Google Scholar), treatment of 5ECER4 cells with Tx led to flattened morphologies reminiscent of trophoblast cells (Figure 1A), induction of trophoblast genes, and downregulation of ES cell genes (Figure 1H). Using trophoblast stem (TS) cell culture conditions (Tanaka et al., 1998Tanaka S. Kunath T. Hadjantonakis A.K. Nagy A. Rossant J. Promotion of trophoblast stem cell proliferation by FGF4.Science. 1998; 282: 2072-2075Crossref PubMed Scopus (999) Google Scholar), TS-like cells could be derived from this line (Figure 1B). These observations are consistent with previous analyses (Niwa et al., 2005Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation.Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (838) Google Scholar) and provide a standard against which to evaluate Tead4 activity in ES cells. We next examined the ability of Tead4 to induce trophoblast fate in ES cells. We established ES cell lines stably expressing a Tx-inducible form of active Tead4 (Tead4VP16ER), which consisted of the Tead4 DNA-binding domain fused to the transcriptional activation domain of herpes simplex virus VP16, followed by the ER domain. After treatment with Tx, multiple independent clones exhibited morphological changes similar to Cdx2 overexpression (Figure 1C). Treated Tead4VP16-expressing clones also expressed trophoblast genes (Figure 1I), and TS-like cells could be derived under TS cell culture conditions (Figure 1D). Thus, constitutively active Tead4 is sufficient to promote trophoblast fate in ES cells. Tead4 is genetically upstream of Cdx2 during TE formation in the embryo (Nishioka et al., 2008Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos.Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (318) Google Scholar, Yagi et al., 2007Yagi R. Kohn M.J. Karavanova I. Kaneko K.J. Vullhorst D. Depamphilis M.L. Buonanno A. Transcription factor TEAD4 specifies the trophectoderm lineage at the beginning of mammalian development.Development. 2007; 134: 3827-3836Crossref PubMed Scopus (339) Google Scholar), suggesting that Tead4 is not required for Cdx2-mediated trophoblast gene expression in ES cells. To test this hypothesis, we established feeder-free Tead4−/− ES cell lines (Nishioka et al., 2008Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos.Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (318) Google Scholar) that stably express Cdx2ER, and we examined their ability to adopt trophoblast fate. After treatment with Tx, multiple independent clones exhibited similar trophoblast-like morphologies (Figure 1E). In addition, trophoblast genes were upregulated, whereas ES genes were downregulated (Figure 1J), suggesting that Tead4 is not required for Cdx2-mediated induction of trophoblast differentiation in ES cells. To examine the requirement for Tead4 in long-term TS-like potential, we cultured these cells under TS conditions. Colonies with TS-like morphology could be derived, but could not be maintained as TS cells (Figure 1F and data not shown), indicating that Cdx2 cannot fully substitute for Tead4 in the trophoblast lineage. As another means by which to examine the epistatic relationship between Tead4 and Cdx2, we established Cdx2−/− ES cells (Niwa et al., 2005Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation.Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (838) Google Scholar) stably expressing Tead4VP16ER, and we examined their ability to adopt trophoblast fate. After treatment with Tx, multiple independent clones also exhibited trophoblast-like morphology (Figure 1G) and trophoblast gene expression (Figure 1K). Tead4 can therefore regulate trophoblast gene expression independently of Cdx2. However, TS-like colonies could not be derived from these cells, suggesting that Cdx2 is essential for proper trophoblast lineage development. Taken together, these observations suggest that Tead4 promotes trophoblast fate through both Cdx2-dependent and -independent pathways, and that Cdx2 is a major mediator of Tead4-dependent changes in trophoblast gene expression. In ES cells, Oct3/4 suppresses Cdx2, and reduction of Oct3/4 leads to upregulation of Cdx2 expression and formation of TS-like cells (Niwa et al., 2000Niwa H. Miyazaki J. Smith A.G. Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells.Nat. Genet. 2000; 24: 372-376Crossref PubMed Scopus (2784) Google Scholar, Niwa et al., 2005Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation.Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (838) Google Scholar). Because Tead4 is required for Cdx2 expression and TE development in vivo (Nishioka et al., 2008Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos.Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (318) Google Scholar, Yagi et al., 2007Yagi R. Kohn M.J. Karavanova I. Kaneko K.J. Vullhorst D. Depamphilis M.L. Buonanno A. Transcription factor TEAD4 specifies the trophectoderm lineage at the beginning of mammalian development.Development. 2007; 134: 3827-3836Crossref PubMed Scopus (339) Google Scholar), we next asked whether Tead4 is required for Cdx2 expression even if Oct3/4 expression levels are reduced. We examined the requirement for Tead4 in inducing Cdx2 expression after siRNA-mediated knockdown of Oct3/4. In control wild-type (EB5) ES cells, siOct3/4 transfection led to reduced expression of Oct3/4 and other ES genes, including Sox2 and Fgf4 (Figure 1L). Knockdown of Oct3/4 also led to increased expression of Cdx2 and other trophoblast genes (Figure 1L). Interestingly, Oct3/4 knockdown in Tead4−/− ES cells (Nishioka et al., 2008Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos.Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (318) Google Scholar) led to essentially the same changes in gene expression as in wild-type ES cells (Figure 1L), indicating that Tead4 is not required for expression of Cdx2 and other trophoblast genes as long as Oct3/4 levels are reduced. However, immediate induction of Cdx2 by Tead4VP16 in ES cells was not accompanied by a clear reduction of Oct3/4 at day 1 (Figure 1I), suggesting that Tead4 can induce Cdx2 expression by overcoming Oct3/4-mediated suppression. Therefore, in the presence of Oct3/4, Tead4 is required to induce Cdx2 expression. These observations suggested that Tead4 can instructively induce Cdx2 expression and trophoblast fate. However, Tead4 is expressed ubiquitously in preimplantation embryos (Nishioka et al., 2008Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos.Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (318) Google Scholar), raising the question as to how its activity is restricted to outer cells of the nascent TE. We hypothesized that Tead4 activity must be regulated along the inside/outside axis. To test this hypothesis, we first examined the ability of variant forms of Tead4 to induce Cdx2 expression in inside cells when overexpressed. During normal development, Cdx2 is initially ubiquitously expressed and becomes progressively downregulated in inside cells and upregulated in outside cells during blastocyst formation (Ralston and Rossant, 2008Ralston A. Rossant J. Cdx2 acts downstream of cell polarization to cell-autonomously promote trophectoderm fate in the early mouse embryo.Dev. Biol. 2008; 313: 614-629Crossref PubMed Scopus (251) Google Scholar). Because levels of Cdx2 are highly variable among embryos and among inside cells of individual embryos during this process (Dietrich and Hiiragi, 2007Dietrich J.E. Hiiragi T. Stochastic patterning in the mouse pre-implantation embryo.Development. 2007; 134: 4219-4231Crossref PubMed Scopus (375) Google Scholar, Ralston and Rossant, 2008Ralston A. Rossant J. Cdx2 acts downstream of cell polarization to cell-autonomously promote trophectoderm fate in the early mouse embryo.Dev. Biol. 2008; 313: 614-629Crossref PubMed Scopus (251) Google Scholar), we examined populations of embryos in which these constructs were ubiquitously overexpressed by RNA injection from the 2-cell stage (Figure 2A). At the 20- to 30-cell stages, embryos were classified into three phenotypic categories, depending on the level of Cdx2 expression detected in inside cells. That is, type I embryos exhibited undetectable levels of Cdx2 in inside cells, type II embryos exhibited low levels of Cdx2 relative to outside cells, and type III embryos exhibited high levels of Cdx2 (comparable to outside cell levels). At this stage, the majority of water or β-globin mRNA-injected embryos exhibited either no Cdx2 (type I) or weak Cdx2 expression (type II) in inside cells, whereas only 8% of embryos on average exhibited strong Cdx2 expression (type III) in a few inside cells (Figures 2C and 2E; see Figure S1 available online). Expression of either full-length Tead4 or a repressor-modified form of Tead4, Tead4EnR (a fusion between the Tead4 containing the DNA-binding domain and the repression domain of Drosophila Engrailed), did not lead to changes in Cdx2 expression (Figures 2B and 2E). In contrast, overexpression of Tead4VP16 led to a significant increase in the number of type III embryos exhibiting elevated Cdx2 expression in inside cells (Figures 2B, 2D, and 2E), consistent with our observations in ES cells. The number of inside cells expressing high levels of Cdx2 also increased after Tead4VP16 overexpression (Figure 2D; Figure S1). Injection of higher doses of Tead4VP16 RNA led to increased lethality (not shown). By contrast, overexpression of an unrelated transcription factor fused to the VP16 activation domain (Foxa2VP16) did not affect Cdx2 expression (Figure 2E). Taken together, these results suggest that the Cdx2-inducing activity of Tead4 is dependent on the presence of an exogenous activation domain. Preimplantation embryos also express Tead1 and Tead2 (Nishioka et al., 2008Nishioka N. Yamamoto S. Kiyonari H. Sato H. Sawada A. Ota M. Nakao K. Sasaki H. Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos.Mech. Dev. 2008; 125: 270-283Crossref PubMed Scopus (318) Google Scholar), and these factors are known to bind similar DNA motifs as Tead4 (Sawada et al., 2008Sawada A. Kiyonari H. Ukita K. Nishioka N. Imuta Y. Sasaki H. Redundant roles of Tead1 and Tead2 in notochord development and the regulation of cell proliferation and survival.Mol. Cell. Biol. 2008; 28: 3177-3189Crossref PubMed Scopus (122) Google Scholar). Interestingly, overexpression of activator-modified Tead1 (Tead1VP16) also increased the frequency of type III embryos (Figure 2E), raising the possibility that other Tead proteins may participate in regulation of Cdx2 expression during embryogenesis. Our analyses of constitutively active Tead4 both in ES cells and in the early embryo suggested that Tead4 activity is regulated along the inside/outside axis of the embryo. Tead proteins are known to act in conjunction with the coactivator protein Yap (Vassilev et al., 2001Vassilev A. Kaneko K.J. Shu H. Zhao Y. DePamphilis M.L. TEAD/TEF transcription factors utilize the activation domain of YAP65, a Src/Yes-associated protein localized in the cytoplasm.Genes Dev. 2001; 15: 1229-1241Crossref PubMed Scopus (472) Google Scholar), whose nuclear localization is regulated by phosphorylation (Zhao et al., 2007Zhao B. Wei X. Li W. Udan R.S. Yang Q. Kim J. Xie J. Ikenoue T. Yu J. Li L. et al.Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control.Genes Dev. 2007; 21: 2747-2761Crossref PubMed Scopus (1767) Google Scholar). Yap mRNA was detected throughout preimplantation development by RT-PCR (Figure 3A), prompting us to examine the localization of Yap protein throughout preimplantation development. During development, nuclear Yap was first detected in embryos at the 4-cell stage, although the signal was weak and variable among embryos and among individual blastomeres (Figure 3B and data not shown). By the early 8-cell stage, nuclear Yap was detected in all blastomeres, even in embryos that had not yet undergone compaction (n = 22) (Figure 3B). After the 8-cell stage, levels of nuclear Yap increased in outside cells up to the 30-cell stage and remained constant thereafter, whereas nuclear Yap decreased in inside cells and Yap appeared to be excluded from the nuclei (Figures 3B and 3D). At the mid/late blastocyst stage, nuclear Yap was restricted to outside cells of the TE and was not detected within cells of the ICM (Figure 3C). Since Cdx2 expression begins after compaction around the 8-cell stage (Ralston and Rossant, 2008Ralston A. Rossant J. Cdx2 acts downstream of cell polarization to cell-autonomously promote trophectoderm fate in the early mouse embryo.Dev. Biol. 2008; 313: 614-629Crossref PubMed Scopus (251) Google Scholar), nuclear localization of Yap appears to precede expression of Cdx2. In addition, restriction of Yap to outside cells appears to precede that of Cdx2, since nuclear Yap was restricted to outside cells from the 16-cell stage onward, whereas Cdx2 is not clearly restricted to outside cells until later stages (Figure 3B) (Dietrich and Hiiragi, 2007Dietrich J.E. Hiiragi T. Stochastic patterning in the mouse pre-implantation embryo.Development. 2007; 134: 4219-4231Crossref PubMed Scopus (375) Google Scholar, Niwa et al., 2005Niwa H. Toyooka Y. Shimosato D. Strumpf D. Takahashi K. Yagi R. Rossant J. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation.Cell. 2005; 123: 917-929Abstract Full Text Full Text PDF PubMed Scopus (838) Google Scholar, Ralston and Rossant, 2008Ralston A. Rossant J. Cdx2 acts downstream of cell polarization to cell-autonomously promote trophectoderm fate in the early mouse embryo.Dev. Biol. 2008; 313: 614-629Crossref PubMed Scopus (251) Google Scholar). Thus, restriction of Yap to outside cell nuclei precedes restriction of Cdx2 expression to outside cells, suggesting that Yap could play a role in Tead4-mediated patterning of Cdx2 expression along the inside/outside axis during blastocyst formation. We next asked whether Yap could induce Cdx2 expression in inside cells of the embryo. Importantly, Yap cooperatively increased transcription induced by Gal4-Tead4C (a fusion protein of the DNA-binding domain of yeast Gal4 and the C-terminal cofactor-binding domain of Tead4) in NIH 3T3 cells (Figure 4B), confirming the ability of Yap to enhance Tead4-mediated transcriptional activity. Next, Yap mRNA was injected into both blastomeres of the 2-cell embryo, and phenotypes were categorized as described above. Overexpression of Yap led to a significant increase in the frequency of type III embryos exhibiting high levels of Cdx2 in inside cells (Figures 4C and 4D). In contrast, neither Yap-ΔTeadBD, lacking the Tead binding domain, nor Yap-ΔAD, lacking the transactivation domain, were able to enhance Tead4-mediated transactivation in NIH 3T3 cells (Figures 4A and 4B), and both constructs failed to increase Cdx2 expression in embryos (Figures 4C and 4D). Thus, Yap activity may depend on interaction with Tead. Overexpression of an unrelated coactivator protein, Ssdp1 (Nishioka et al., 2005Nishioka N. Nagano S. Nakayama R. Kiyonari H. Ijiri T. Taniguchi K. Shawlot W. Hayashizaki Y. Westphal H. Behringer R.R. et al.Ssdp1 regulates head morphogenesis of mouse embryos by activating the Lim1-Ldb1 complex.Development. 2005; 132: 2535-2546Crossref PubMed Scopus (54) Google Scholar), had no effect on Tead4-mediated transcription in NIH 3T3 cells, or on Cdx2 expression in embryos (Figures 4B and 4D), confirming the specificity of our observations. Finally, to examine whether Yap can confer Cdx2-inducing ability on unmodified Tead4 (lacking VP16), we examined the ability of unmodified Tead4 to induce Cdx2 expression in the presence of Yap. Injection of either full-length Tead4 mRNA or low-dose (5 ng/μl) Yap mRNA alone had no effect on Cdx2 expression, whereas their coinjection significantly increased Cdx2 expression (Figures 4C and 4D). Taken together, our observations suggest that Yap can induce Cdx2 expression cooperatively with Tead4. Although Yap is sufficient to upregulate Cdx2 in inside cells, Yap−/− mutant embryos exhibit normal TE development (Morin-Kensicki et al., 2006Morin-Kensicki E.M. Boone B.N. Howell M. Stonebraker J.R. Teed J. Alb J.G. Magnuson T.R. O'Neal W. Milgram S.L. Defects in yolk sac vasculogenesis, chorioallantoic fusion, and embryonic axis elongation in mice with targeted disruption of Yap65.Mol. Cell. Biol. 2006; 26: 77-87Crossref PubMed Scopus (276) Google Scholar). This observation suggested that a Yap-related protein could compensate for the absence of Yap during early development. We therefore examined the Yap-related protein Wwtr1 (TAZ) (Mahoney et al., 2005Mahoney Jr., W.M. Hong J.H. Yaffe M.B. Farrance I.K. The transcriptional co-activator TAZ interacts differentially with transcriptional enhancer factor-1 (TEF-1) family members.Biochem. J. 2005; 388: 217-225Crossref PubMed Scopus (159) Google Scholar). Wwtr1 was detected in preimplantation embryos; high levels of Wwtr1 protein were detected in outside cell nuclei (Figures 3A and 3E), whereas low, but detectable, levels of Wwtr1 were detected in inside cell nuclei. Importantly, overex" @default.
• W2004102023 created "2016-06-24" @default.
• W2004102023 creator A5011048453 @default.
• W2004102023 creator A5011985445 @default.
• W2004102023 creator A5033183436 @default.
• W2004102023 creator A5034949085 @default.
• W2004102023 creator A5035181971 @default.
• W2004102023 creator A5035867151 @default.
• W2004102023 creator A5041029568 @default.
• W2004102023 creator A5043769666 @default.
• W2004102023 creator A5048858084 @default.
• W2004102023 creator A5056496717 @default.
• W2004102023 creator A5058178004 @default.
• W2004102023 creator A5059427033 @default.
• W2004102023 creator A5060578424 @default.
• W2004102023 creator A5060872339 @default.
• W2004102023 creator A5064949815 @default.
• W2004102023 creator A5074594797 @default.
• W2004102023 creator A5080516181 @default.
• W2004102023 creator A5081180990 @default.
• W2004102023 date "2009-03-01" @default.
• W2004102023 modified "2023-10-15" @default.
• W2004102023 title "The Hippo Signaling Pathway Components Lats and Yap Pattern Tead4 Activity to Distinguish Mouse Trophectoderm from Inner Cell Mass" @default.
• W2004102023 cites W1531523780 @default.
• W2004102023 cites W1608762409 @default.
• W2004102023 cites W1975751233 @default.
• W2004102023 cites W1978746441 @default.
• W2004102023 cites W1984282087 @default.
• W2004102023 cites W1986676916 @default.
• W2004102023 cites W1994144105 @default.
• W2004102023 cites W1996420367 @default.
• W2004102023 cites W2000225283 @default.
• W2004102023 cites W2006499356 @default.
• W2004102023 cites W2007455356 @default.
• W2004102023 cites W2011108752 @default.
• W2004102023 cites W2011818472 @default.
• W2004102023 cites W2011946635 @default.
• W2004102023 cites W2012305943 @default.
• W2004102023 cites W2030433725 @default.
• W2004102023 cites W2032942434 @default.
• W2004102023 cites W2034820114 @default.
• W2004102023 cites W2046985565 @default.
• W2004102023 cites W2050552243 @default.
• W2004102023 cites W2053482584 @default.
• W2004102023 cites W2055378633 @default.
• W2004102023 cites W2060020667 @default.
• W2004102023 cites W2065786458 @default.
• W2004102023 cites W2069375339 @default.
• W2004102023 cites W2076823362 @default.
• W2004102023 cites W2079346360 @default.
• W2004102023 cites W2083870204 @default.
• W2004102023 cites W2085197940 @default.
• W2004102023 cites W2088863538 @default.
• W2004102023 cites W2090465993 @default.
• W2004102023 cites W2091054780 @default.
• W2004102023 cites W2092081014 @default.
• W2004102023 cites W2095573728 @default.
• W2004102023 cites W2097192405 @default.
• W2004102023 cites W2099105062 @default.
• W2004102023 cites W2099709490 @default.
• W2004102023 cites W2111622605 @default.
• W2004102023 cites W2118611175 @default.
• W2004102023 cites W2122864540 @default.
• W2004102023 cites W2123829995 @default.
• W2004102023 cites W2128323764 @default.
• W2004102023 cites W2128718933 @default.
• W2004102023 cites W2130746708 @default.
• W2004102023 cites W2133811885 @default.
• W2004102023 cites W2134386050 @default.
• W2004102023 cites W2135726634 @default.
• W2004102023 cites W2138312252 @default.
• W2004102023 cites W2141595533 @default.
• W2004102023 cites W2143539586 @default.
• W2004102023 cites W2157193997 @default.
• W2004102023 cites W2158342928 @default.
• W2004102023 cites W2159196011 @default.
• W2004102023 cites W2164384065 @default.
• W2004102023 cites W2167978765 @default.
• W2004102023 doi "https://doi.org/10.1016/j.devcel.2009.02.003" @default.
• W2004102023 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/19289085" @default.
• W2004102023 hasPublicationYear "2009" @default.
• W2004102023 type Work @default.
• W2004102023 sameAs 2004102023 @default.
• W2004102023 citedByCount "860" @default.
• W2004102023 countsByYear W20041020232012 @default.
• W2004102023 countsByYear W20041020232013 @default.
• W2004102023 countsByYear W20041020232014 @default.
• W2004102023 countsByYear W20041020232015 @default.
• W2004102023 countsByYear W20041020232016 @default.
• W2004102023 countsByYear W20041020232017 @default.
• W2004102023 countsByYear W20041020232018 @default.
• W2004102023 countsByYear W20041020232019 @default.
• W2004102023 countsByYear W20041020232020 @default.
• W2004102023 countsByYear W20041020232021 @default.
• W2004102023 countsByYear W20041020232022 @default.
• W2004102023 countsByYear W20041020232023 @default.
• W2004102023 crossrefType "journal-article" @default.
• W2004102023 hasAuthorship W2004102023A5011048453 @default.

• next