MCM10 mediates RECQ4 association with MCM2-7 helicase complex during DNA replication.
Mutations in RECQ4, a member of the RecQ family of DNA helicases, have been linked to the progeroid disease Rothmund-Thomson Syndrome. Attempts to understand the complex phenotypes observed in recq4-deficient cells suggest a potential involvement in DNA repair and replication, yet the molecular basis of the function of RECQ4 in these processes remains unknown. Here, we report the identification of a highly purified chromatin-bound RECQ4 complex from human cell extracts. We found that essential replisome factors MCM10, MCM2-7 helicase, CDC45 and GINS are the primary interaction partner proteins of human RECQ4. Importantly, complex formation and the association of RECQ4 with the replication origin are cell-cycle regulated. Furthermore, we show that MCM10 is essential for the integrity of the RECQ4-MCM replicative helicase complex. MCM10 interacts directly with RECQ4 and regulates its DNA unwinding activity, and that this interaction may be modulated by cyclin-dependent kinase phosphorylation. Thus, these studies show that RECQ4 is an integral component of the MCM replicative helicase complex participating in DNA replication in human cells.
Xu
Xiaohua
X
Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT 06520-8040, USA.
Rochette
Patrick J
PJ
Feyissa
Eminet A
EA
Su
Tina V
TV
Liu
Yilun
Y
D000818:Animals; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000235:genetics; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D064111:Minichromosome Maintenance Complex Component 2; D064168:Minichromosome Maintenance Complex Component 7; D064110:Minichromosome Maintenance Proteins; D009687:Nuclear Proteins / Q000378:metabolism*; D053484:RecQ Helicases / Q000302:isolation & purification / Q000378:metabolism*; D018741:Replication Origin
D018797:Cell Cycle Proteins; D002843:Chromatin; D004268:DNA-Binding Proteins; C489613:MCM10 protein, human; D009687:Nuclear Proteins; D004247:DNA; C506731:RECQL4 protein, human; C084859:MCM2 protein, human; C101624:MCM7 protein, human; D064111:Minichromosome Maintenance Complex Component 2; D064168:Minichromosome Maintenance Complex Component 7; D064110:Minichromosome Maintenance Proteins; D053484:RecQ Helicases
Nature. 2002 Feb 7;415(6872):651-5
11807498
Nature. 2005 Mar 31;434(7033):598-604
15800615
Mol Cell. 2002 Feb;9(2):233-40
11864598
Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8580-4
18562274
Genes Dev. 2008 Oct 15;22(20):2737-42
18923071
Genes Dev. 2007 Dec 15;21(24):3331-41
18079179
Methods. 2009 May;48(1):72-9
19245838
Cell Mol Life Sci. 2002 Aug;59(8):1317-26
12363035
Mol Cell Biol. 1998 Oct;18(10):6102-9
9742127
Cell. 2005 Jun 17;121(6):887-98
15960976
Mol Cell Biol. 2006 Jul;26(13):4843-52
16782873
Hum Mol Genet. 2003 Sep 15;12(18):2293-9
12915449
Proc Natl Acad Sci U S A. 2007 Sep 18;104(38):14929-34
17846426
Proc Natl Acad Sci U S A. 2008 Jul 1;105(26):8956-61
18579778
Hum Mol Genet. 2004 Oct 15;13(20):2421-30
15317757
Nature. 2007 Jan 18;445(7125):281-5
17167417
Cell Div. 2007 Jun 05;2:16
17547773
Nature. 2007 Jan 18;445(7125):328-32
17167415
Front Biosci. 1999 Dec 01;4:D859-68
10577395
Genes Dev. 2000 Apr 15;14(8):913-26
10783164
Biogerontology. 2009 Jun;10(3):235-52
19083132
Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2334-9
12604790
EMBO J. 2009 Mar 4;28(5):568-77
19177149
J Mol Biol. 2007 Feb 9;366(1):36-52
17141802
Mol Cell. 2006 Oct 6;24(1):101-13
17018296
EMBO J. 2006 May 3;25(9):1987-96
16619031
Cell. 2005 Sep 23;122(6):915-26
16153703
Annu Rev Genet. 2007;41:237-80
17630848
Hum Mol Genet. 2005 Mar 15;14(6):813-25
15703196
Curr Biol. 1999 Sep 9;9(17):975-8
10508584
Nihon Yakurigaku Zasshi. 2002 Apr;119(4):219-26
11979727
Int J Biochem Cell Biol. 2002 Nov;34(11):1496-501
12200042