title: The C-terminal domain of Brd2 is important for chromatin interaction and regulation of transcription and alternative splicing.
abstract: Brd2 is a member of the bromodomain extra terminal (BET) protein family, which consists of four chromatin-interacting proteins that regulate gene expression. Each BET protein contains two N-terminal bromodomains, which recognize acetylated histones, and the C-terminal protein-protein interaction domain. Using a genome-wide screen, we identify 1450 genes whose transcription is regulated by Brd2. In addition, almost 290 genes change their alternative splicing pattern upon Brd2 depletion. Brd2 is specifically localized at promoters of target genes, and our data show that Brd2 interaction with chromatin cannot be explained solely by histone acetylation. Using coimmunoprecipitation and live-cell imaging, we show that the C-terminal part is crucial for Brd2 association with chromatin. Live-cell microscopy also allows us to map the average binding time of Brd2 to chromatin and quantify the contributions of individual Brd2 domains to the interaction with chromatin. Finally, we show that bromodomains and the C-terminal domain are equally important for transcription and splicing regulation, which correlates with the role of these domains in Brd2 binding to chromatin.
authors.affiliation: Department of RNA Biology, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague 142 20, Czech Republic Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague 142 20, Czech Republic Max Planck Institute for Molecular Cell Biology and Genetics, Dresden 01307, Germany J. Heyrovsky Institute of Physical Chemistry, Academy of Sciences of the Czech Republic, Prague 182 23, Czech Republic.
authors.forename: Jarmila
authors.initials: J
authors.lastname: Hnilicová
authors.forename: Samira
authors.initials: S
authors.lastname: Hozeifi
authors.forename: Eva
authors.initials: E
authors.lastname: Stejskalová
authors.forename: Eva
authors.initials: E
authors.lastname: Dušková
authors.forename: Ina
authors.initials: I
authors.lastname: Poser
authors.forename: Jana
authors.initials: J
authors.lastname: Humpolíčková
authors.forename: Martin
authors.initials: M
authors.lastname: Hof
authors.forename: David
authors.initials: D
authors.lastname: Staněk
mesh_terms: D017398:Alternative Splicing; D002843:Chromatin / Q000378:metabolism*; D005786:Gene Expression Regulation; D015894:Genome, Human; D006367:HeLa Cells; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D018715:Microscopy, Video; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014157:Transcription Factors; D014158:Transcription, Genetic; D000092468:Bromodomain Containing Proteins
chemical_list: D002843:Chromatin; D006657:Histones; D017346:Protein Serine-Threonine Kinases; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; C098415:BRD2 protein, human; D000092468:Bromodomain Containing Proteins
references.citation: Nat Rev Cancer. 2012 Jun 22;12(7):465-77
references.pmid: 22722403
references.citation: Nucleus. 2011 May-Jun;2(3):182-8
references.pmid: 21818411
references.citation: Nat Struct Mol Biol. 2010 Sep;17(9):1114-23
references.pmid: 20711188
references.citation: Nature. 2013 Jun 13;498(7453):241-5
references.pmid: 23739326
references.citation: Wiley Interdiscip Rev RNA. 2013 Jan-Feb;4(1):77-91
references.pmid: 23074139
references.citation: J Cell Sci. 2012 Aug 1;125(Pt 15):3671-80
references.pmid: 22595521
references.citation: FEBS Lett. 2010 Sep 24;584(18):3901-8
references.pmid: 20709061
references.citation: Cell. 2013 Mar 14;152(6):1252-69
references.pmid: 23498935
references.citation: Mol Cell. 2005 Aug 19;19(4):535-45
references.pmid: 16109377
references.citation: Cell. 2012 Jul 6;150(1):12-27
references.pmid: 22770212
references.citation: Nat Struct Mol Biol. 2011 Jul 26;18(9):977-83
references.pmid: 21792193
references.citation: Science. 2009 Jul 3;325(5936):90-3
references.pmid: 19574390
references.citation: Biochim Biophys Acta. 2009 May;1789(5):413-21
references.pmid: 19362612
references.citation: Biophys J. 2004 Jun;86(6):3473-95
references.pmid: 15189848
references.citation: J Microsc. 2008 Jan;229(Pt 1):78-91
references.pmid: 18173647
references.citation: BMC Struct Biol. 2007 Sep 12;7:57
references.pmid: 17848202
references.citation: BMC Bioinformatics. 2009 Feb 03;10:48
references.pmid: 19192299
references.citation: Genome Biol. 2012 Aug 16;13(8):R68
references.pmid: 22897906
references.citation: Biochem J. 2005 Apr 1;387(Pt 1):257-69
references.pmid: 15548137
references.citation: Nat Rev Cancer. 2009 Jan;9(1):40-55
references.pmid: 19078974
references.citation: Br J Cancer. 2011 Jan 4;104(1):155-65
references.pmid: 21081927
references.citation: Mol Cell Biochem. 2007 Jan;294(1-2):45-54
references.pmid: 17111193
references.citation: Proc Natl Acad Sci U S A. 2011 May 31;108(22):E159-68
references.pmid: 21536911
references.citation: Science. 2010 Feb 19;327(5968):996-1000
references.pmid: 20133523
references.citation: Nucleic Acids Res. 2012 Aug;40(15):7162-75
references.pmid: 22570411
references.citation: Cell. 2009 Jul 10;138(1):129-45
references.pmid: 19596240
references.citation: Cell. 2010 Sep 17;142(6):967-80
references.pmid: 20850016
references.citation: J Clin Invest. 2011 May;121(5):1905-16
references.pmid: 21490398
references.citation: PLoS One. 2011;6(8):e23656
references.pmid: 21887291
references.citation: Mol Cell Biol. 2011 Jul;31(13):2641-52
references.pmid: 21555454
references.citation: PLoS Genet. 2009 Oct;5(10):e1000682
references.pmid: 19834536
references.citation: Nat Genet. 2000 May;25(1):25-9
references.pmid: 10802651
references.citation: Science. 2002 Oct 11;298(5592):416-9
references.pmid: 12376702
references.citation: Mol Cell. 2004 Jan 16;13(1):33-43
references.pmid: 14731392
references.citation: Cell Rep. 2012 Jul 26;2(1):62-8
references.pmid: 22840397
references.citation: Mol Cell Biochem. 2006 Nov;292(1-2):205-12
references.pmid: 16786191
references.citation: J Immunol. 2007 Oct 15;179(8):5462-73
references.pmid: 17911633
references.citation: Dev Dyn. 2009 Apr;238(4):908-17
references.pmid: 19301389
references.citation: Cell. 2011 Jan 7;144(1):16-26
references.pmid: 21215366
references.citation: PLoS One. 2011;6(7):e21800
references.pmid: 21789182
references.citation: Microsc Res Tech. 2009 Apr;72(4):323-32
references.pmid: 19067357
references.citation: Mol Cell. 2005 Aug 19;19(4):523-34
references.pmid: 16109376
references.citation: Cell. 2008 Apr 4;133(1):192.e1
references.pmid: 18394998
references.citation: FEBS Lett. 2012 Aug 14;586(17):2692-704
references.pmid: 22710155
references.citation: Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11456-60
references.pmid: 9326631
references.citation: J Cell Biol. 2010 Oct 4;191(1):75-86
references.pmid: 20921136
references.citation: J Biol Chem. 2004 Jul 9;279(28):29075-84
references.pmid: 15126509
references.citation: Proc Natl Acad Sci U S A. 2012 May 1;109(18):6927-32
references.pmid: 22509028
references.citation: J Biol Chem. 2003 Oct 31;278(44):43034-40
references.pmid: 12939267
references.citation: Biophys J. 2005 Aug;89(2):1317-27
references.pmid: 15908582
references.citation: Proc Natl Acad Sci U S A. 2009 Mar 17;106(11):4325-30
references.pmid: 19251664
references.citation: Cell Growth Differ. 2000 Aug;11(8):417-24
references.pmid: 10965846
references.citation: Nature. 2010 Dec 23;468(7327):1119-23
references.pmid: 21068722
references.citation: Biochem J. 2009 Dec 14;425(1):71-83
references.pmid: 19883376
references.citation: Trends Cell Biol. 2011 Jun;21(6):328-35
references.pmid: 21530266
references.citation: Mol Biol Cell. 2009 Dec;20(23):4899-909
references.pmid: 19812244
references.citation: Acta Biomater. 2011 Dec;7(12):4195-203
references.pmid: 21801861
references.citation: Trends Pharmacol Sci. 2012 Mar;33(3):146-53
references.pmid: 22277300
references.citation: PLoS One. 2011 Feb 02;6(2):e16727
references.pmid: 21311748
references.citation: RNA. 2013 May;19(5):627-38
references.pmid: 23525800
references.citation: J Natl Cancer Inst. 2010 Jun 16;102(12):866-80
references.pmid: 20484105
references.citation: Nat Methods. 2008 May;5(5):409-15
references.pmid: 18391959
references.citation: Curr Opin Cell Biol. 2005 Jun;17(3):262-8
references.pmid: 15901495
references.citation: Epilepsia. 2007 Apr;48(4):706-12
references.pmid: 17437413
references.citation: Nat Genet. 2013 Aug;45(8):852-9
references.pmid: 23793028
references.citation: Mol Cell. 2000 Aug;6(2):307-16
references.pmid: 10983978
references.citation: J Proteome Res. 2006 Mar;5(3):502-11
references.pmid: 16512664
references.citation: Chem Biol. 2011 Apr 22;18(4):495-507
references.pmid: 21513886
references.citation: Mol Cell. 2008 Apr 11;30(1):51-60
references.pmid: 18406326
references.citation: Mol Cell Biol. 2011 Jul;31(13):2632-40
references.pmid: 21555453
references.citation: Nat Cell Biol. 2011 Oct 09;13(11):1295-304
references.pmid: 21983563
references.citation: Mol Endocrinol. 2002 Aug;16(8):1727-37
references.pmid: 12145330
references.citation: Mol Cell. 2007 Nov 30;28(4):665-76
references.pmid: 18042460
references.citation: Cell. 2012 Mar 30;149(1):214-31
references.pmid: 22464331
references.citation: Am J Hum Genet. 2003 Aug;73(2):261-70
references.pmid: 12830434
references.citation: Proc Natl Acad Sci U S A. 2003 Jul 22;100(15):8758-63
references.pmid: 12840145
references.citation: Oncogene. 1998 Mar 26;16(12):1513-23
references.pmid: 9569018
references.citation: Proc Natl Acad Sci U S A. 2002 Jun 11;99(12):8185-90
references.pmid: 12060763
references.citation: J Biol Chem. 2002 Nov 8;277(45):43110-4
references.pmid: 12221105
references.citation: Biophys J. 2005 May;88(5):3601-14
references.pmid: 15722439