title: Cyclin D1 promotes cell cycle progression through enhancing NDR1/2 kinase activity independent of cyclin-dependent kinase 4.
abstract: Cyclin/cyclin-dependent kinases (Cdks) are critical protein kinases in regulating cell cycle progression. Among them, cyclin D1/Cdk4 exerts its function mainly in the G1 phase. By using the tandem affinity purification tag approach, we identified a set of proteins interacting with Cdk4, including NDR1/2. Interestingly, confirming the interactions between NDR1/2 and cyclin D1/Cdk4, we observed that NDR1/2 interacted with cyclin D1 independent of Cdk4, but NDR1/2 and cyclin D1/Cdk4 did not phosphorylate each other. In addition, we found that NDR1/2 did not affect the kinase activity of cyclin D1/Cdk4 upon phosphorylation of GST-Rb. However, cyclin D1 but not Cdk4 promoted the kinase activity of NDR1/2. We also demonstrated that cyclin D1 K112E, which could not bind Cdk4, enhanced the kinase activity of NDR1/2. To test whether cyclin D1 promotes G1/S transition though enhancing NDR1/2 kinase activity, we performed flow cytometry analysis using cyclin D1 and cyclin D1 K112E Tet-On inducible cell lines. The data show that both cyclin D1 and cyclin D1 K112E promoted G1/S transition. Importantly, knockdown of NDR1/2 almost completely abolished the function of cyclin D1 K112E in promoting G1/S transition. Consistently, we found that the protein level of p21 was reduced in cells overexpressing cyclin D1 K112E but not when NDR1/2 was knocked down. Taken together, these results reveal a novel function of cyclin D1 in promoting cell cycle progression by enhancing NDR kinase activity independent of Cdk4.
authors.affiliation: From the Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101 and.
authors.forename: Zhaoyang
authors.initials: Z
authors.lastname: Du
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Tong
authors.forename: Xin
authors.initials: X
authors.lastname: Ye
mesh_terms: D002453:Cell Cycle; D019938:Cyclin D1 / Q000378:metabolism*; D051358:Cyclin-Dependent Kinase 4 / Q000378:metabolism*; D005434:Flow Cytometry; D015971:Gene Expression Regulation, Enzymologic; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D025941:Protein Interaction Mapping; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D034622:RNA Interference
chemical_list: C530104:CCND1 protein, human; D019938:Cyclin D1; D017346:Protein Serine-Threonine Kinases; C491614:STK38 protein, human; C496014:STK38L protein, human; C495900:CDK4 protein, human; D051358:Cyclin-Dependent Kinase 4
keywords: Cell Cycle; Cyclins; Protein Phosphorylation; Protein-Protein Interactions; Serine/Threonine Protein Kinase
references.citation: J Biol Chem. 2006 May 19;281(20):13915-21
references.pmid: 16488889
references.citation: J Biol Chem. 2004 May 28;279(22):23806-12
references.pmid: 15037617
references.citation: Science. 1993 Dec 24;262(5142):2050-4
references.pmid: 8266103
references.citation: J Biol Chem. 2002 Mar 15;277(11):8847-53
references.pmid: 11788592
references.citation: Mol Cell Biol. 2011 Apr;31(7):1382-95
references.pmid: 21262772
references.citation: Nature. 2011 Jun 08;474(7350):230-4
references.pmid: 21654808
references.citation: Cancer Cell. 2011 Nov 15;20(5):620-34
references.pmid: 22094256
references.citation: Mol Cell Biol. 1996 Nov;16(11):6457-67
references.pmid: 8887674
references.citation: Genes Dev. 1997 Apr 1;11(7):847-62
references.pmid: 9106657
references.citation: Cell. 1993 Nov 19;75(4):805-16
references.pmid: 8242751
references.citation: J Biol Chem. 2001 May 18;276(20):16840-7
references.pmid: 11279133
references.citation: J Biol Chem. 2003 Jul 11;278(28):25752-7
references.pmid: 12730199
references.citation: Mol Cell Biol. 2010 Sep;30(18):4507-20
references.pmid: 20624913
references.citation: Mol Cell. 2007 Feb 23;25(4):625-34
references.pmid: 17317633
references.citation: Nature. 2004 Jul 8;430(6996):226-31
references.pmid: 15241418
references.citation: J Biol Chem. 2004 Aug 20;279(34):35228-35
references.pmid: 15197186
references.citation: J Biol Chem. 2009 Mar 20;284(12):7875-88
references.pmid: 19150984
references.citation: Curr Biol. 2009 Apr 28;19(8):675-81
references.pmid: 19327996
references.citation: Mol Cell Biol. 1998 Mar;18(3):1590-600
references.pmid: 9488476
references.citation: Genes Dev. 1999 Jun 15;13(12):1501-12
references.pmid: 10385618
references.citation: Curr Biol. 2008 Dec 9;18(23):1889-95
references.pmid: 19062280
references.citation: Cell. 1997 Feb 7;88(3):405-15
references.pmid: 9039267
references.citation: Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):709-13
references.pmid: 8290586
references.citation: J Biol Chem. 2003 Feb 28;278(9):6710-8
references.pmid: 12493777
references.citation: Mol Cell Biol. 2011 Sep;31(17):3603-15
references.pmid: 21730291