title: Activation of a human chromosomal replication origin by protein tethering.
abstract: The specification of mammalian chromosomal replication origins is incompletely understood. To analyze the assembly and activation of prereplicative complexes (pre-RCs), we tested the effects of tethered binding of chromatin acetyltransferases and replication proteins on chromosomal c-myc origin deletion mutants containing a GAL4-binding cassette. GAL4(DBD) (DNA binding domain) fusions with Orc2, Cdt1, E2F1 or HBO1 coordinated the recruitment of the Mcm7 helicase subunit, the DNA unwinding element (DUE)-binding protein DUE-B and the minichromosome maintenance (MCM) helicase activator Cdc45 to the replicator, and restored origin activity. In contrast, replication protein binding and origin activity were not stimulated by fusion protein binding in the absence of flanking c-myc DNA. Substitution of the GAL4-binding site for the c-myc replicator DUE allowed Orc2 and Mcm7 binding, but eliminated origin activity, indicating that the DUE is essential for pre-RC activation. Additionally, tethering of DUE-B was not sufficient to recruit Cdc45 or activate pre-RCs formed in the absence of a DUE. These results show directly in a chromosomal background that chromatin acetylation, Orc2 or Cdt1 suffice to recruit all downstream replication initiation activities to a prospective origin, and that chromosomal origin activity requires singular DNA sequences.
authors: Department of Biochemistry and Molecular Biology, Boonshoft School of Medicine, Wright State University, Dayton, OH 45435, USA.
authors: Xiaomi
authors: X
authors: Chen
authors: Guoqi
authors: G
authors: Liu
authors: Michael
authors: M
authors: Leffak
mesh_terms: D018797:Cell Cycle Proteins / Q000378:metabolism; D002877:Chromosomes, Human / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D050687:E2F1 Transcription Factor / Q000378:metabolism; D016259:Genes, myc; D006367:HeLa Cells; D051548:Histone Acetyltransferases / Q000378:metabolism; D006801:Humans; D064168:Minichromosome Maintenance Complex Component 7; D009687:Nuclear Proteins / Q000378:metabolism; D051738:Origin Recognition Complex / Q000378:metabolism; D018741:Replication Origin
chemical_list: C467199:CDT1 protein, human; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D050687:E2F1 Transcription Factor; D009687:Nuclear Proteins; D051738:Origin Recognition Complex; D051548:Histone Acetyltransferases; C101624:MCM7 protein, human; D064168:Minichromosome Maintenance Complex Component 7
references: Nucleic Acids Res. 2011 Mar;39(6):2103-15
references: 21062817
references: Genes Dev. 2010 Jan 15;24(2):183-94
references: 20080954
references: Genome Res. 2010 Feb;20(2):201-11
references: 19996087
references: Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2656-61
references: 10077566
references: Nat Struct Mol Biol. 2011 Apr;18(4):471-7
references: 21378962
references: Nucleic Acids Res. 2005 Jan 13;33(1):325-36
references: 15653633
references: Curr Biol. 2009 Mar 24;19(6):530-5
references: 19285403
references: Chromosoma. 2007 Apr;116(2):197-214
references: 17219175
references: J Cell Sci. 2012 Feb 15;125(Pt 4):965-72
references: 22421364
references: Mol Cell Biol. 2008 Jan;28(1):140-53
references: 17954561
references: Mol Cell Biol. 1999 Aug;19(8):5685-95
references: 10409757
references: J Cell Biol. 2006 Jun 5;173(5):673-83
references: 16754955
references: Mol Cell Biol. 2012 Aug;32(15):3107-20
references: 22645314
references: Nature. 2004 Jul 15;430(6997):372-6
references: 15254542
references: EMBO J. 2004 Feb 25;23(4):897-907
references: 14765124
references: Genome Res. 2006 May;16(5):595-605
references: 16606705
references: Mol Cell Biol. 2006 Nov;26(21):7977-90
references: 16940174
references: J Biol Chem. 2004 Oct 1;279(40):41686-94
references: 15284237
references: J Biol Chem. 2007 Apr 6;282(14):10441-8
references: 17264083
references: Proc Natl Acad Sci U S A. 2009 Dec 1;106(48):20240-5
references: 19910535
references: J Biol Chem. 2012 Sep 28;287(40):33412-23
references: 22872635
references: Genes Dev. 2008 Oct 1;22(19):2633-8
references: 18832067
references: Proc Natl Acad Sci U S A. 1989 May;86(10):3594-8
references: 2524830
references: Genes Dev. 2011 Mar 15;25(6):621-33
references: 21406556
references: Curr Opin Cell Biol. 2006 Jun;18(3):231-9
references: 16650748
references: J Biol Chem. 2004 Mar 5;279(10):9625-33
references: 14672932
references: J Biol Chem. 2011 Apr 1;286(13):11855-64
references: 21282109
references: J Biol Chem. 2007 Apr 20;282(16):11705-14
references: 17314092
references: J Biol Chem. 1999 Aug 13;274(33):23027-34
references: 10438470
references: Mol Cell Biol. 2012 May;32(9):1618-32
references: 22354993
references: Science. 2007 Feb 2;315(5812):653-5
references: 17272723
references: Curr Biol. 2011 Jul 12;21(13):1152-7
references: 21700459
references: Genome Biol. 2012 Apr 24;13(4):R27
references: 22531001
references: Mol Cell Biol. 2001 Feb;21(4):1098-110
references: 11158297
references: Curr Biol. 2000 Feb 24;10(4):195-205
references: 10704410
references: Nature. 2000 Apr 6;404(6778):622-5
references: 10766247
references: Mol Cell Biol. 2004 Dec;24(23):10193-207
references: 15542830
references: Eur J Biochem. 1999 Nov;265(3):936-43
references: 10518787
references: Nat Cell Biol. 2006 Jan;8(1):84-90
references: 16362051
references: Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12309-14
references: 8901577
references: Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):6914-8
references: 8346196
references: FEMS Yeast Res. 2007 Dec;7(8):1256-62
references: 17825064
references: J Cell Sci. 2002 Apr 1;115(Pt 7):1435-40
references: 11896191
references: Mol Syst Biol. 2010 Aug 24;6:404
references: 20739926
references: Proc Natl Acad Sci U S A. 2009 Sep 15;106(37):15628-32
references: 19805216
references: Mol Cell Biol. 2006 Jul;26(14):5270-83
references: 16809765
references: Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13589-94
references: 11717425
references: Genes Dev. 2003 Aug 1;17(15):1894-908
references: 12897055
references: Mol Cell. 2003 Apr;11(4):997-1008
references: 12718885
references: FEMS Yeast Res. 2012 Jun;12(4):486-90
references: 22380713
references: J Mol Med (Berl). 2002 Aug;80(8):463-74
references: 12185447
references: EMBO J. 1996 Dec 2;15(23):6671-9
references: 8978693
references: Mol Cell. 2008 Jul 25;31(2):190-200
references: 18657502
references: Genes Dev. 2005 Dec 1;19(23):2827-36
references: 16322558
references: J Biol Chem. 2001 Jul 13;276(28):26666-73
references: 11323433
references: Cell. 2011 Sep 16;146(6):931-41
references: 21925316
references: J Biol Chem. 2001 Nov 30;276(48):44905-11
references: 11555648
references: Curr Biol. 2011 Aug 23;21(16):R638-40
references: 21855008
references: Cell Cycle. 2010 Nov 1;9(21):4351-63
references: 20980834
references: J Biol Chem. 2001 May 4;276(18):15397-408
references: 11278932
references: Genes Dev. 2010 Apr 15;24(8):748-53
references: 20351051
references: Cancer Res. 2007 Nov 15;67(22):10899-909
references: 18006835
references: J Biol Chem. 2005 Apr 1;280(13):13071-83
references: 15653697
references: Nucleic Acids Res. 2009 Jan;37(1):60-9
references: 19010964
references: J Cell Biol. 2004 May 24;165(4):473-82
references: 15159417
references: Mol Cell Biol. 2004 Aug;24(16):7140-50
references: 15282313
references: J Biol Chem. 2009 Apr 24;284(17):11446-53
references: 19276081
references: Genes Dev. 2007 Nov 15;21(22):2897-907
references: 18006685
references: Mol Cell. 2010 Jan 15;37(1):57-66
references: 20129055
references: Cell. 2010 Feb 5;140(3):349-59
references: 20116089
references: Mol Cell Biol. 2001 Dec;21(23):8095-103
references: 11689699
references: J Biol Chem. 2008 Sep 12;283(37):25356-25363
references: 18617514
references: Nat Cell Biol. 2010 May;12(5):484-91
references: 20383140
references: Mol Cell Biol. 2010 Mar;30(6):1495-507
references: 20065034
references: Mol Cell Biol. 2003 Mar;23(5):1832-42
references: 12589000
references: Nucleic Acids Res. 2005 Feb 01;33(2):765-75
references: 15687385
references: Mol Cell Biol. 2001 Sep;21(17):6006-16
references: 11486038
references: J Biol Chem. 2008 Oct 31;283(44):30216-24
references: 18723846
references: Genes Dev. 1999 Sep 15;13(18):2360-4
references: 10500092
references: Cell Cycle. 2012 Feb 15;11(4):658-67
references: 22373526
references: Genes Dev. 1999 Apr 1;13(7):827-40
references: 10197983
references: Genes Dev. 2005 Jan 1;19(1):114-26
references: 15598982
references: Mol Cell Biol. 2004 May;24(10):4546-56
references: 15121871
references: Curr Opin Cell Biol. 2009 Dec;21(6):771-7
references: 19767190
references: Mol Cell Biol. 2004 May;24(10):4138-50
references: 15121836
references: Genome Res. 2008 Nov;18(11):1763-77
references: 18836037
references: Mol Cell. 2010 Jan 29;37(2):247-58
references: 20122406
references: Mol Cell Biol. 2001 Oct;21(20):6820-32
references: 11564866
references: J Biol Chem. 2004 Aug 20;279(34):35879-89
references: 15190069
references: Nucleic Acids Res. 2001 Feb 1;29(3):809-17
references: 11160905
references: Curr Biol. 2005 Jan 26;15(2):146-53
references: 15668171
references: Genes Dev. 2007 Mar 1;21(5):497-518
references: 17344412
references: Oncogene. 1994 Feb;9(2):405-15
references: 8290253
references: Nat Chem Biol. 2010 Sep;6(9):652-9
references: 20676085
references: Annu Rev Biochem. 2010;79:89-130
references: 20373915
references: Mol Cell Biol. 2007 Nov;27(22):7828-38
references: 17846122
references: Mol Cell. 2001 Jan;7(1):21-30
references: 11172708