title: Regulation of the mouse Treacher Collins syndrome homolog (Tcof1) promoter through differential repression of constitutive expression.
abstract: Treacher Collins syndrome is an autosomal-dominant mandibulofacial dysostosis caused by haploinsufficiency of the TCOF1 gene product treacle. Mouse Tcof1 protein is approximately 61% identical and 71% similar to treacle, and heterozygous knockout of Tcof1 causes craniofacial malformation. Tcof1 expression is high in developing neural crest, but much lower in other tissues. To investigate this dual regulation, highly conserved regions upstream of TCOF1 homologs were tested through deletion and mutation reporter assays, and conserved predicted transcription factor binding sites were assessed through chromatin binding studies. Assays were performed in mouse P19 embryonic carcinoma cells and in HEK293 cells to determine differential activation in cell types at different stages of differentiation. Binding of Cebpb, Zfp161, and Sp1 transcription factors was specific to the Tcof1 regulatory region in P19 cells. The Zfp161 binding site demonstrated P19 cell-specific repression, while the Sp1/Sp3 candidate site demonstrated HEK293 cell-specific activation. Moreover, presence of c-myb and Zfp161 transcripts was specific to P19 cells. A minimal promoter fragment from -253 to +43 bp directs constitutive expression in both cell types, and dual regulation of Tcof1 appears to be through differential repression of this minimal promoter. The CpG island at the transcription start site remains unmethylated in P19 cells, 11.5 dpc mouse embryonic tissue, and adult mouse ear, which supports constitutive activation of the Tcof1 promoter.
authors.affiliation: Department of Human and Molecular Genetics, Virginia Commonwealth University School of Medicine, Richmond, Virginia 23298-0033, USA. khshows@vcu.edu
authors.forename: Kathryn H
authors.initials: KH
authors.lastname: Shows
authors.forename: Rita
authors.initials: R
authors.lastname: Shiang
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D018899:CpG Islands; D019175:DNA Methylation; D005786:Gene Expression Regulation; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D010750:Phosphoproteins; C105442:TCOF1 protein, human; C491352:Tcof1 protein, mouse; D014157:Transcription Factors
references.citation: FEMS Microbiol Lett. 1999 May 15;174(2):247-50
references.pmid: 10339815
references.citation: Development. 2001 Aug;128(16):3049-60
references.pmid: 11688555
references.citation: Nucleic Acids Res. 1993 Aug 11;21(16):3767-75
references.pmid: 8367294
references.citation: Am J Hum Genet. 1997 Mar;60(3):515-24
references.pmid: 9042910
references.citation: Hum Mol Genet. 1996 Oct;5(10):1533-8
references.pmid: 8894686
references.citation: Proc Natl Acad Sci U S A. 2003 Jan 21;100(2):532-7
references.pmid: 12511599
references.citation: Hum Mutat. 2000 Oct;16(4):315-22
references.pmid: 11013442
references.citation: Hum Mol Genet. 1997 May;6(5):727-37
references.pmid: 9158147
references.citation: Am J Med Genet A. 2003 Dec 1;123A(2):169-71
references.pmid: 14598341
references.citation: Adv Otorhinolaryngol. 2000;56:53-9
references.pmid: 10868214
references.citation: Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):3110-5
references.pmid: 9096354
references.citation: Cell Mol Life Sci. 2005 Nov;62(21):2516-25
references.pmid: 16231084
references.citation: Mol Cell Biol. 1987 Mar;7(3):1101-10
references.pmid: 3561410
references.citation: Biochem Biophys Res Commun. 1997 Sep 8;238(1):1-6
references.pmid: 9299440
references.citation: Dev Dyn. 2004 Apr;229(4):907-14
references.pmid: 15042714
references.citation: Biochem J. 2002 Aug 1;365(Pt 3):561-75
references.pmid: 12006103
references.citation: Nucleic Acids Res. 1997 Mar 15;25(6):1108-16
references.pmid: 9092617
references.citation: Proc Natl Acad Sci U S A. 2002 Jan 22;99(2):602-7
references.pmid: 11805317
references.citation: Nucleic Acids Res. 1996 Mar 15;24(6):1091-8
references.pmid: 8604343
references.citation: Blood Cells Mol Dis. 2003 Sep-Oct;31(2):192-200
references.pmid: 12972026
references.citation: Mol Biol Cell. 2000 Sep;11(9):3061-71
references.pmid: 10982400
references.citation: J Virol. 2004 Jul;78(14):7590-601
references.pmid: 15220434
references.citation: Hum Mol Genet. 1998 Nov;7(12):1947-52
references.pmid: 9811939
references.citation: Biochem Biophys Res Commun. 1999 Feb 16;255(2):528-34
references.pmid: 10049742
references.citation: Birth Defects Res C Embryo Today. 2005 Sep;75(3):200-12
references.pmid: 16187326
references.citation: Gene. 2005 Oct 10;359:44-52
references.pmid: 16102917
references.citation: Science. 2002 Mar 22;295(5563):2258-61
references.pmid: 11910109
references.citation: Hum Mol Genet. 2005 Jul 15;14(14):2035-43
references.pmid: 15930015
references.citation: Int J Dev Biol. 1993 Mar;37(1):135-40
references.pmid: 8507558
references.citation: Hum Mol Genet. 1998 Oct;7(11):1795-800
references.pmid: 9736782
references.citation: Science. 2004 Dec 24;306(5705):2255-7
references.pmid: 15618518
references.citation: Proc Natl Acad Sci U S A. 2006 Sep 5;103(36):13403-8
references.pmid: 16938878
references.citation: Biotechniques. 2007 Nov;43(5):683-93
references.pmid: 18072598
references.citation: Mamm Genome. 2006 Feb;17(2):168-77
references.pmid: 16465596
references.citation: Birth Defects Res A Clin Mol Teratol. 2004 Dec;70(12):912-26
references.pmid: 15578713
references.citation: Hum Mol Genet. 2000 Jun 12;9(10):1473-80
references.pmid: 10888597
references.citation: Curr Protoc Bioinformatics. 2008 Mar;Chapter 2:Unit 2.6
references.pmid: 18428685
references.citation: Biochem Biophys Res Commun. 2005 Feb 11;327(2):615-27
references.pmid: 15629158
references.citation: Genome Res. 2002 Jun;12(6):996-1006
references.pmid: 12045153
references.citation: Mol Cell Biol. 2000 Oct;20(20):7572-82
references.pmid: 11003653
references.citation: Int J Biochem Cell Biol. 1997 Dec;29(12):1313-23
references.pmid: 9570130
references.citation: J Med Genet. 2002 Jul;39(7):493-5
references.pmid: 12114482
references.citation: Biochem Biophys Res Commun. 1999 Mar 24;256(3):573-8
references.pmid: 10080939
references.citation: Proc Natl Acad Sci U S A. 2004 Jul 20;101(29):10709-14
references.pmid: 15249688
references.citation: J Appl Genet. 2002;43(2):223-33
references.pmid: 12080178
references.citation: Biochem Biophys Res Commun. 2004 Dec 3;325(1):124-32
references.pmid: 15522210
references.citation: Hum Mutat. 2005 May;25(5):429-34
references.pmid: 15832313
references.citation: Genome Res. 1997 Mar;7(3):223-34
references.pmid: 9074926
references.citation: Eur J Hum Genet. 2004 Nov;12(11):879-90
references.pmid: 15340364
references.citation: Gene. 2004 Mar 17;328:49-57
references.pmid: 15019983
references.citation: Biochim Biophys Acta. 1997 May 2;1352(1):23-6
references.pmid: 9177479
references.citation: J Biol Chem. 2004 Nov 5;279(45):46826-34
references.pmid: 15326190
references.citation: J Biol Chem. 2003 Sep 5;278(36):34309-19
references.pmid: 12777385
references.citation: Nat Genet. 1996 Feb;12(2):130-6
references.pmid: 8563749
references.citation: Mamm Genome. 2001 Aug;12(8):622-9
references.pmid: 11471057