MiR-322/424 and -503 are induced during muscle differentiation and promote cell cycle quiescence and differentiation by down-regulation of Cdc25A.
Induction of a G1 phase cell cycle arrest, caused primarily by the inhibition of cyclin-dependent-kinase 2 (cdk2), is a critical step in the differentiation of myoblasts into myotubes. Here, we report that two microRNAs, miR-322/424 and miR-503, are induced and promote cdk2 inhibition during myogenesis. These microRNAs down-regulate Cdc25A, the phosphatase responsible for removing inhibitory phosphorylation of cdk2, both in myoblasts differentiating into myotubes and in nonmuscle cells. Cdc25A is down-regulated during muscle differentiation by multiple pathways: action of these two microRNAs, proteasomal degradation of Cdc25A protein and transcriptional repression. Overexpression of Cdc25A or of cdk2 with mutations on T14 and Y15 (cdk2-AF), so that it cannot be inhibited by phosphorylation, decreases differentiation and differentiation-induced cell cycle quiescence. Introduction of miR-322/424 and miR-503 in heterologous cancer cells induces G1 arrest, which is also attenuated by overexpression of the cdk2-AF mutant. Until now Cdc25A and the inhibitory phosphorylation on T14 and Y15 of cdk2 have only been implicated in the intra-S phase checkpoint pathway after DNA damage. Our results reveal an unexpected role of Cdc25A down-regulation and the inhibitory phosphorylation of cdk2 T14 and Y15 in cell cycle quiescence during muscle differentiation and implicate two muscle differentiation-induced microRNAs in the process.
Sarkar
Sukumar
S
Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.
Dey
Bijan K
BK
Dutta
Anindya
A
D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D045744:Cell Line, Tumor; D051357:Cyclin-Dependent Kinase 2 / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000502:physiology*; D020687:cdc25 Phosphatases / Q000235:genetics / Q000378:metabolism*
C582672:MIRN322 microRNA, mouse; C554214:MIRN424 microrna, human; D035683:MicroRNAs; D051357:Cyclin-Dependent Kinase 2; C517004:CDC25A protein, human; D020687:cdc25 Phosphatases
Exp Cell Res. 2004 Nov 1;300(2):365-78
15475001
Cancer Res. 1995 Nov 15;55(22):5187-90
7585571
Gene. 2000 Apr 18;247(1-2):1-15
10773440
J Cell Biol. 1996 Feb;132(4):657-66
8647896
Proc Natl Acad Sci U S A. 2009 Aug 11;106(32):13383-7
19666532
J Cell Biol. 2006 Aug 28;174(5):677-87
16923828
EMBO J. 2002 Sep 16;21(18):4875-84
12234927
Nat Genet. 2006 Feb;38(2):228-33
16380711
Oncogene. 2006 Aug 28;25(38):5210-9
16936739
Proc Natl Acad Sci U S A. 2005 Dec 27;102(52):18986-91
16357195
Proc Natl Acad Sci U S A. 2005 May 3;102(18):6279-84
15845771
Mol Biol Cell. 2009 Apr;20(8):2186-95
19244340
Science. 2005 Jul 8;309(5732):310-1
15919954
Cancer Res. 2009 Oct 15;69(20):8157-65
19826040
Genes Dev. 2007 Jan 1;21(1):71-84
17210790
Stem Cells. 2009 Jul;27(7):1524-8
19544458
Curr Biol. 2003 Oct 14;13(20):1775-85
14561402
Cell. 2007 Apr 20;129(2):303-17
17397913
Nat Cell Biol. 2006 Mar;8(3):278-84
16489342
Anticancer Agents Med Chem. 2008 Dec;8(8):825-31
19075564
J Clin Invest. 2009 Aug;119(8):2366-78
19620785
Genes Dev. 2008 Dec 1;22(23):3242-54
19015276
J Biol Chem. 2009 Oct 23;284(43):29596-604
19710019
Mol Cell. 2009 Sep 11;35(5):610-25
19748357
Nat Genet. 2003 Sep;35(1):25-31
12923533
Nature. 1977 Dec 22-29;270(5639):725-7
563524
J Cell Sci. 2009 Jan 1;122(Pt 1):13-20
19092056
Science. 2000 May 26;288(5470):1425-9
10827953
EMBO J. 1992 Nov;11(11):3995-4005
1396589
Curr Opin Genet Dev. 2006 Oct;16(5):533-9
16919443
Prog Cell Cycle Res. 2000;4:107-14
10740819
Nature. 1995 Mar 9;374(6518):131-4
7877684
Annu Rev Pathol. 2009;4:199-227
18817506
Cancer Res. 2007 Aug 15;67(16):7713-22
17699775
Proc Natl Acad Sci U S A. 2007 Dec 11;104(50):19849-54
18056638
Cancer Res. 2007 Jul 15;67(14):6605-11
17638870
Exp Biol Med (Maywood). 2008 Feb;233(2):131-8
18222968
EMBO J. 1991 Feb;10(2):305-16
1846803
Mol Cell Biol. 2000 Oct;20(19):7059-67
10982822
Genes Dev. 2003 Dec 15;17(24):3062-74
14681206
J Biol Chem. 2008 Apr 11;283(15):9836-43
18281287
Dev Biol. 2007 Nov 15;311(2):359-68
17936265
Oncogene. 2006 Aug 28;25(38):5244-9
16936743
Genes Dev. 2001 Oct 15;15(20):2654-9
11641272
Cancer Cell. 2008 Nov 4;14(5):369-81
18977326
Genes Dev. 2004 Nov 15;18(22):2699-711
15545627
Am J Physiol Cell Physiol. 2003 Nov;285(5):C1019-27
12826599
Curr Opin Cell Biol. 2009 Jun;21(3):461-9
19278845
Nat Cell Biol. 2009 Oct;11(10):1247-53
19734889
J Neurooncol. 2001 Feb;51(3):205-18
11407593
Nature. 2003 Nov 6;426(6962):87-91
14603323