{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T19:40:42Z","timestamp":1790278842991,"version":"4.1.0"},"reference-count":55,"publisher":"The Company of Biologists","issue":"6","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2013,3,15]]},"abstract":"<jats:p>Cleft palate is one of the most common human birth defects and is associated with multiple genetic and environmental risk factors. Although mutations in the genes encoding transforming growth factor beta (TGF\u03b2) signaling molecules and interferon regulatory factor 6 (Irf6) have been identified as genetic risk factors for cleft palate, little is known about the relationship between TGF\u03b2 signaling and IRF6 activity during palate formation. Here, we show that TGF\u03b2 signaling regulates expression of Irf6 and the fate of the medial edge epithelium (MEE) during palatal fusion in mice. Haploinsufficiency of Irf6 in mice with basal epithelial-specific deletion of the TGF\u03b2 signaling mediator Smad4 (Smad4fl\/fl;K14-Cre;Irf6+\/R84C) results in compromised p21 expression and MEE persistence, similar to observations in Tgfbr2fl\/fl;K14-Cre mice, although the secondary palate of Irf6+\/R84C and Smad4fl\/fl;K14-Cre mice form normally. Furthermore, Smad4fl\/fl;K14-Cre;Irf6+\/R84C mice show extra digits that are consistent with abnormal toe and nail phenotypes in individuals with Van der Woude and popliteal pterygium syndromes, suggesting that the TGF\u03b2\/SMAD4\/IRF6 signaling cascade might be a well-conserved mechanism in regulating multiple organogenesis. Strikingly, overexpression of Irf6 rescued p21 expression and MEE degeneration in Tgfbr2fl\/fl;K14-Cre mice. Thus, IRF6 and SMAD4 synergistically regulate the fate of the MEE, and TGF\u03b2-mediated Irf6 activity is responsible for MEE degeneration during palatal fusion in mice.<\/jats:p>","DOI":"10.1242\/dev.089615","type":"journal-article","created":{"date-parts":[[2013,2,13]],"date-time":"2013-02-13T22:19:33Z","timestamp":1360793973000},"page":"1220-1230","source":"Crossref","is-referenced-by-count":83,"title":["<i>Smad4<\/i>\n                    -\n                    <i>Irf6<\/i>\n                    genetic interaction and TGF\u03b2-mediated IRF6 signaling cascade are crucial for palatal fusion in mice"],"prefix":"10.1242","volume":"140","author":[{"given":"Jun-ichi","family":"Iwata","sequence":"first","affiliation":[{"name":"Center for Craniofacial Molecular Biology, Ostrow School of Dentistry, University of Southern California, Los Angeles, CA 90033, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akiko","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Center for Craniofacial Molecular Biology, Ostrow School of Dentistry, University of Southern California, Los Angeles, CA 90033, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard C.","family":"Pelikan","sequence":"additional","affiliation":[{"name":"Center for Craniofacial Molecular Biology, Ostrow School of Dentistry, University of Southern California, Los Angeles, CA 90033, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thach-Vu","family":"Ho","sequence":"additional","affiliation":[{"name":"Center for Craniofacial Molecular Biology, Ostrow School of Dentistry, University of Southern California, Los Angeles, CA 90033, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pedro A.","family":"Sanchez-Lara","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA."},{"name":"Division of Medical Genetics, Children\u2019s Hospital Los Angeles, Los Angeles, CA 90033, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark","family":"Urata","sequence":"additional","affiliation":[{"name":"Center for Craniofacial Molecular Biology, Ostrow School of Dentistry, University of Southern California, Los Angeles, CA 90033, USA."},{"name":"Division of Plastic Surgery, Children\u2019s Hospital Los Angeles, Los Angeles, CA 90033, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael J.","family":"Dixon","sequence":"additional","affiliation":[{"name":"Faculty of Life Sciences and Dental School, University of Manchester, Manchester, UK."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Chai","sequence":"additional","affiliation":[{"name":"Center for Craniofacial Molecular Biology, Ostrow School of Dentistry, University of Southern California, Los Angeles, CA 90033, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","reference":[{"key":"2021042607321096900_R1","doi-asserted-by":"crossref","first-page":"1165","DOI":"10.1002\/ajmg.a.33968","article-title":"SMAD4 mutation segregating in a family with juvenile polyposis, aortopathy, and mitral valve dysfunction","volume":"155","author":"Andrabi","year":"2011","journal-title":"Am. J. Med. Genet. A."},{"key":"2021042607321096900_R2","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1146\/annurev.cellbio.14.1.167","article-title":"Transcriptional control of muscle development by myocyte enhancer factor-2 (MEF2) proteins","volume":"14","author":"Black","year":"1998","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"2021042607321096900_R3","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1093\/jb\/mvs053","article-title":"A half-site of the p53-binding site on the keratin 14 promoter is specifically activated by p63","volume":"152","author":"Cai","year":"2012","journal-title":"J. Biochem."},{"key":"2021042607321096900_R4","doi-asserted-by":"crossref","first-page":"1959","DOI":"10.1038\/sj.onc.1202111","article-title":"Suppression of transforming growth factor-beta-induced apoptosis through a phosphatidylinositol 3-kinase\/Akt-dependent pathway","volume":"17","author":"Chen","year":"1998","journal-title":"Oncogene"},{"key":"2021042607321096900_R5","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1242\/dev.00907","article-title":"Death is the major fate of medial edge epithelial cells and the cause of basal lamina degradation during palatogenesis","volume":"131","author":"Cuervo","year":"2004","journal-title":"Development"},{"key":"2021042607321096900_R6","doi-asserted-by":"crossref","first-page":"6397","DOI":"10.1073\/pnas.090099297","article-title":"Transforming growth factor beta -inducible independent binding of SMAD to the Smad7 promoter","volume":"97","author":"Denissova","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2021042607321096900_R7","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1038\/nrg2933","article-title":"Cleft lip and palate: understanding genetic and environmental influences","volume":"12","author":"Dixon","year":"2011","journal-title":"Nat. Rev. Genet."},{"key":"2021042607321096900_R8","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1038\/ng0492-45","article-title":"Definition of a consensus binding site for p53","volume":"1","author":"el-Deiry","year":"1992","journal-title":"Nat. Genet."},{"key":"2021042607321096900_R9","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1002\/dvdy.22786","article-title":"MCS9.7 enhancer activity is highly, but not completely, associated with expression of Irf6 and p63","volume":"241","author":"Fakhouri","year":"2012","journal-title":"Dev. Dyn."},{"key":"2021042607321096900_R10","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1016\/j.devcel.2011.08.005","article-title":"A conserved Pbx-Wnt-p63-Irf6 regulatory module controls face morphogenesis by promoting epithelial apoptosis","volume":"21","author":"Ferretti","year":"2011","journal-title":"Dev. Cell"},{"key":"2021042607321096900_R11","doi-asserted-by":"crossref","first-page":"5028","DOI":"10.1093\/emboj\/18.18.5028","article-title":"Crystal structure of an IRF-DNA complex reveals novel DNA recognition and cooperative binding to a tandem repeat of core sequences","volume":"18","author":"Fujii","year":"1999","journal-title":"EMBO J."},{"key":"2021042607321096900_R12","doi-asserted-by":"crossref","first-page":"2524","DOI":"10.1093\/hmg\/dds071","article-title":"Tbx1 regulates oral epithelial adhesion and palatal development","volume":"21","author":"Funato","year":"2012","journal-title":"Hum. Mol. Genet."},{"key":"2021042607321096900_R13","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1136\/jmg.2006.041517","article-title":"SMAD4 mutations found in unselected HHT patients","volume":"43","author":"Gallione","year":"2006","journal-title":"J. Med. Genet."},{"key":"2021042607321096900_R14","doi-asserted-by":"crossref","first-page":"4531","DOI":"10.1038\/sj.onc.1203806","article-title":"TGFbeta1 represses proliferation of pancreatic carcinoma cells which correlates with Smad4-independent inhibition of ERK activation","volume":"19","author":"Giehl","year":"2000","journal-title":"Oncogene"},{"key":"2021042607321096900_R15","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/S1050-1738(00)00039-6","article-title":"Regulation of MEF2 by p38 MAPK and its implication in cardiomyocyte biology","volume":"10","author":"Han","year":"2000","journal-title":"Trends Cardiovasc. Med."},{"key":"2021042607321096900_R16","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1016\/j.ydbio.2010.12.021","article-title":"Epithelial Wnt\/\u03b2-catenin signaling regulates palatal shelf fusion through regulation of Tgf\u03b23 expression","volume":"350","author":"He","year":"2011","journal-title":"Dev. Biol."},{"key":"2021042607321096900_R17","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1093\/emboj\/18.5.1345","article-title":"TGF-beta induces fibronectin synthesis through a c-Jun N-terminal kinase-dependent, Smad4-independent pathway","volume":"18","author":"Hocevar","year":"1999","journal-title":"EMBO J."},{"key":"2021042607321096900_R18","doi-asserted-by":"crossref","first-page":"35381","DOI":"10.1074\/jbc.274.50.35381","article-title":"The MEK pathway is required for stimulation of p21(WAF1\/CIP1) by transforming growth factor-beta","volume":"274","author":"Hu","year":"1999","journal-title":"J. Biol. Chem."},{"key":"2021042607321096900_R19","doi-asserted-by":"crossref","first-page":"1335","DOI":"10.1038\/ng1903","article-title":"Abnormal skin, limb and craniofacial morphogenesis in mice deficient for interferon regulatory factor 6 (Irf6)","volume":"38","author":"Ingraham","year":"2006","journal-title":"Nat. Genet."},{"key":"2021042607321096900_R20","doi-asserted-by":"crossref","first-page":"5269","DOI":"10.1242\/dev.00708","article-title":"Conditional inactivation of Tgfbr2 in cranial neural crest causes cleft palate and calvaria defects","volume":"130","author":"Ito","year":"2003","journal-title":"Development"},{"key":"2021042607321096900_R21","doi-asserted-by":"crossref","first-page":"4035","DOI":"10.1074\/jbc.M512283200","article-title":"Excess peroxisomes are degraded by autophagic machinery in mammals","volume":"281","author":"Iwata","year":"2006","journal-title":"J. Biol. Chem."},{"key":"2021042607321096900_R22","doi-asserted-by":"crossref","first-page":"4975","DOI":"10.1074\/jbc.M109.035105","article-title":"Transforming growth factor-beta regulates basal transcriptional regulatory machinery to control cell proliferation and differentiation in cranial neural crest-derived osteoprogenitor cells","volume":"285","author":"Iwata","year":"2010","journal-title":"J. Biol. Chem."},{"key":"2021042607321096900_R23","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1111\/j.1601-0825.2011.01806.x","article-title":"The mechanism of TGF-\u03b2 signaling during palate development","volume":"17","author":"Iwata","year":"2011","journal-title":"Oral Dis."},{"key":"2021042607321096900_R24","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1172\/JCI61498","article-title":"Modulation of noncanonical TGF-\u03b2 signaling prevents cleft palate in Tgfbr2 mutant mice","volume":"122","author":"Iwata","year":"2012","journal-title":"J. Clin. Invest."},{"key":"2021042607321096900_R25","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1016\/j.tcb.2009.05.008","article-title":"New regulatory mechanisms of TGF-beta receptor function","volume":"19","author":"Kang","year":"2009","journal-title":"Trends Cell Biol."},{"key":"2021042607321096900_R26","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1136\/jmg.2004.019646","article-title":"A genome-wide scan of non-syndromic cleft palate only (CPO) in Finnish multiplex families","volume":"42","author":"Koillinen","year":"2005","journal-title":"J. Med. Genet."},{"key":"2021042607321096900_R27","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1038\/ng985","article-title":"Mutations in IRF6 cause Van der Woude and popliteal pterygium syndromes","volume":"32","author":"Kondo","year":"2002","journal-title":"Nat. Genet."},{"key":"2021042607321096900_R28","doi-asserted-by":"crossref","first-page":"e1001065","DOI":"10.1371\/journal.pgen.1001065","article-title":"Genome-wide profiling of p63 DNA-binding sites identifies an element that regulates gene expression during limb development in the 7q21 SHFM1 locus","volume":"6","author":"Kouwenhoven","year":"2010","journal-title":"PLoS Genet."},{"key":"2021042607321096900_R29","doi-asserted-by":"crossref","first-page":"2947","DOI":"10.1093\/bioinformatics\/btm404","article-title":"Clustal W and Clustal X version 2.0","volume":"23","author":"Larkin","year":"2007","journal-title":"Bioinformatics"},{"key":"2021042607321096900_R30","doi-asserted-by":"crossref","first-page":"1553","DOI":"10.1242\/dev.02325","article-title":"p63 regulates multiple signalling pathways required for ectodermal organogenesis and differentiation","volume":"133","author":"Laurikkala","year":"2006","journal-title":"Development"},{"key":"2021042607321096900_R31","doi-asserted-by":"crossref","first-page":"e45441","DOI":"10.1371\/journal.pone.0045441","article-title":"Interaction between IRF6 and TGFA genes contribute to the risk of nonsyndromic cleft lip\/palate","volume":"7","author":"Letra","year":"2012","journal-title":"PLoS ONE"},{"key":"2021042607321096900_R32","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1093\/hmg\/ddn381","article-title":"Missense mutations that cause Van der Woude syndrome and popliteal pterygium syndrome affect the DNA-binding and transcriptional activation functions of IRF6","volume":"18","author":"Little","year":"2009","journal-title":"Hum. Mol. Genet."},{"key":"2021042607321096900_R33","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1038\/nrm3434","article-title":"TGF\u03b2 signalling in context","volume":"13","author":"Massagu\u00e9","year":"2012","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2021042607321096900_R34","doi-asserted-by":"crossref","first-page":"1570","DOI":"10.1172\/JCI40267","article-title":"A regulatory feedback loop involving p63 and IRF6 links the pathogenesis of 2 genetically different human ectodermal dysplasias","volume":"120","author":"Moretti","year":"2010","journal-title":"J. Clin. Invest."},{"key":"2021042607321096900_R35","doi-asserted-by":"crossref","first-page":"1773","DOI":"10.1016\/S0140-6736(09)60695-4","article-title":"Cleft lip and palate","volume":"374","author":"Mossey","year":"2009","journal-title":"Lancet"},{"key":"2021042607321096900_R36","doi-asserted-by":"crossref","first-page":"4171","DOI":"10.1093\/hmg\/ddp368","article-title":"Tbx22null mice have a submucous cleft palate due to reduced palatal bone formation and also display ankyloglossia and choanal atresia phenotypes","volume":"18","author":"Pauws","year":"2009","journal-title":"Hum. Mol. Genet."},{"key":"2021042607321096900_R37","doi-asserted-by":"crossref","first-page":"7363","DOI":"10.1038\/sj.onc.1210561","article-title":"p63 consensus DNA-binding site: identification, analysis and application into a p63MH algorithm","volume":"26","author":"Perez","year":"2007","journal-title":"Oncogene"},{"key":"2021042607321096900_R38","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1038\/ng1894","article-title":"Irf6 is a key determinant of the keratinocyte proliferation-differentiation switch","volume":"38","author":"Richardson","year":"2006","journal-title":"Nat. Genet."},{"key":"2021042607321096900_R39","doi-asserted-by":"crossref","first-page":"2632","DOI":"10.1093\/hmg\/ddp201","article-title":"Integration of IRF6 and Jagged2 signalling is essential for controlling palatal adhesion and fusion competence","volume":"18","author":"Richardson","year":"2009","journal-title":"Hum. Mol. Genet."},{"key":"2021042607321096900_R40","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/j.biocel.2007.09.006","article-title":"How the Smads regulate transcription","volume":"40","author":"Ross","year":"2008","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"2021042607321096900_R41","doi-asserted-by":"crossref","first-page":"970","DOI":"10.1038\/nrm2297","article-title":"TGFbeta-SMAD signal transduction: molecular specificity and functional flexibility","volume":"8","author":"Schmierer","year":"2007","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2021042607321096900_R42","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1016\/S0092-8674(03)00432-X","article-title":"Mechanisms of TGF-beta signaling from cell membrane to the nucleus","volume":"113","author":"Shi","year":"2003","journal-title":"Cell"},{"key":"2021042607321096900_R43","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1038\/ncb1780","article-title":"The type I TGF-beta receptor engages TRAF6 to activate TAK1 in a receptor kinase-independent manner","volume":"10","author":"Sorrentino","year":"2008","journal-title":"Nat. Cell Biol."},{"key":"2021042607321096900_R44","doi-asserted-by":"crossref","first-page":"e46","DOI":"10.1136\/jmg.2005.032235","article-title":"Significant association between IRF6 820G-&gt;A and non-syndromic cleft lip with or without cleft palate in the Thai population","volume":"42","author":"Srichomthong","year":"2005","journal-title":"J. Med. Genet."},{"key":"2021042607321096900_R45","doi-asserted-by":"crossref","first-page":"1561","DOI":"10.1172\/JCI40266","article-title":"Cooperation between the transcription factors p63 and IRF6 is essential to prevent cleft palate in mice","volume":"120","author":"Thomason","year":"2010","journal-title":"J. Clin. Invest."},{"key":"2021042607321096900_R46","doi-asserted-by":"crossref","first-page":"52087","DOI":"10.1074\/jbc.M404802200","article-title":"Cell-specific activation of the atrial natriuretic factor promoter by PITX2 and MEF2A","volume":"279","author":"Toro","year":"2004","journal-title":"J. Biol. Chem."},{"key":"2021042607321096900_R47","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.ydbio.2009.09.028","article-title":"Signaling integration in the rugae growth zone directs sequential SHH signaling center formation during the rostral outgrowth of the palate","volume":"336","author":"Welsh","year":"2009","journal-title":"Dev. Biol."},{"key":"2021042607321096900_R48","doi-asserted-by":"crossref","first-page":"2264","DOI":"10.1128\/MCB.23.7.2264-2276.2003","article-title":"The Delta Np63 alpha phosphoprotein binds the p21 and 14-3-3 sigma promoters in vivo and has transcriptional repressor activity that is reduced by Hay-Wells syndrome-derived mutations","volume":"23","author":"Westfall","year":"2003","journal-title":"Mol. Cell. Biol."},{"key":"2021042607321096900_R49","first-page":"331","article-title":"An update on the aetiology of orofacial clefts","volume":"10","author":"Wong","year":"2004","journal-title":"Hong Kong Med. J."},{"key":"2021042607321096900_R50","doi-asserted-by":"crossref","first-page":"238","DOI":"10.1016\/j.ydbio.2006.05.014","article-title":"Cell autonomous requirement for Tgfbr2 in the disappearance of medial edge epithelium during palatal fusion","volume":"297","author":"Xu","year":"2006","journal-title":"Dev. Biol."},{"key":"2021042607321096900_R51","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1016\/j.devcel.2008.06.004","article-title":"Ectodermal Smad4 and p38 MAPK are functionally redundant in mediating TGF-beta\/BMP signaling during tooth and palate development","volume":"15","author":"Xu","year":"2008","journal-title":"Dev. Cell"},{"key":"2021042607321096900_R52","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1016\/j.molcel.2008.09.002","article-title":"TRAF6 mediates Smad-independent activation of JNK and p38 by TGF-beta","volume":"31","author":"Yamashita","year":"2008","journal-title":"Mol. Cell"},{"key":"2021042607321096900_R53","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1002\/dvg.20372","article-title":"Tissue-specific expression of Cre recombinase from the Tgfb3 locus","volume":"46","author":"Yang","year":"2008","journal-title":"Genesis"},{"key":"2021042607321096900_R54","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1016\/S1097-2765(00)80061-1","article-title":"Human Smad3 and Smad4 are sequence-specific transcription activators","volume":"1","author":"Zawel","year":"1998","journal-title":"Mol. Cell"},{"key":"2021042607321096900_R55","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1038\/cr.2008.328","article-title":"Non-Smad pathways in TGF-beta signaling","volume":"19","author":"Zhang","year":"2009","journal-title":"Cell Res."}],"container-title":["Development"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/journals.biologists.com\/dev\/article-pdf\/140\/6\/1220\/1321371\/1220.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/journals.biologists.com\/dev\/article-pdf\/140\/6\/1220\/1321371\/1220.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,4,26]],"date-time":"2021-04-26T07:43:13Z","timestamp":1619422993000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.biologists.com\/dev\/article\/140\/6\/1220\/46053\/Smad4-Irf6-genetic-interaction-and-TGF-mediated"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,3,15]]},"references-count":55,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2013,3,15]]}},"URL":"https:\/\/doi.org\/10.1242\/dev.089615","relation":{},"ISSN":["1477-9129","0950-1991"],"issn-type":[{"value":"1477-9129","type":"electronic"},{"value":"0950-1991","type":"print"}],"subject":[],"published":{"date-parts":[[2013,3,15]]}}}