A dominant-negative cyclin D1 mutant prevents nuclear import of cyclin-dependent kinase 4 (CDK4) and its phosphorylation by CDK-activating kinase - PubMed (original) (raw)
A dominant-negative cyclin D1 mutant prevents nuclear import of cyclin-dependent kinase 4 (CDK4) and its phosphorylation by CDK-activating kinase
J A Diehl et al. Mol Cell Biol. 1997 Dec.
Abstract
Cyclins contain two characteristic cyclin folds, each consisting of five alpha-helical bundles, which are connected to one another by a short linker peptide. The first repeat makes direct contact with cyclin-dependent kinase (CDK) subunits in assembled holoenzyme complexes, whereas the second does not contribute directly to the CDK interface. Although threonine 156 in mouse cyclin D1 is predicted to lie at the carboxyl terminus of the linker peptide that separates the two cyclin folds and is buried within the cyclin subunit, mutation of this residue to alanine has profound effects on the behavior of the derived cyclin D1-CDK4 complexes. CDK4 in complexes with mutant cyclin D1 (T156A or T156E but not T156S) is not phosphorylated by recombinant CDK-activating kinase (CAK) in vitro, fails to undergo activating T-loop phosphorylation in vivo, and remains catalytically inactive and unable to phosphorylate the retinoblastoma protein. Moreover, when it is ectopically overexpressed in mammalian cells, cyclin D1 (T156A) assembles with CDK4 in the cytoplasm but is not imported into the cell nucleus. CAK phosphorylation is not required for nuclear transport of cyclin D1-CDK4 complexes, because complexes containing wild-type cyclin D1 and a CDK4 (T172A) mutant lacking the CAK phosphorylation site are efficiently imported. In contrast, enforced overexpression of the CDK inhibitor p21Cip1 together with mutant cyclin D1 (T156A)-CDK4 complexes enhanced their nuclear localization. These results suggest that cyclin D1 (T156A or T156E) forms abortive complexes with CDK4 that prevent recognition by CAK and by other cellular factors that are required for their nuclear localization. These properties enable ectopically overexpressed cyclin D1 (T156A), or a more stable T156A/T286A double mutant that is resistant to ubiquitination, to compete with endogenous cyclin D1 in mammalian cells, thereby mobilizing CDK4 into cytoplasmic, catalytically inactive complexes and dominantly inhibiting the ability of transfected NIH 3T3 fibroblasts to enter S phase.
Similar articles
- Regulation of cyclin D-dependent kinase 4 (cdk4) by cdk4-activating kinase.
Kato JY, Matsuoka M, Strom DK, Sherr CJ. Kato JY, et al. Mol Cell Biol. 1994 Apr;14(4):2713-21. doi: 10.1128/mcb.14.4.2713-2721.1994. Mol Cell Biol. 1994. PMID: 8139570 Free PMC article. - Critical role of cyclin D1 nuclear import in cardiomyocyte proliferation.
Tamamori-Adachi M, Ito H, Sumrejkanchanakij P, Adachi S, Hiroe M, Shimizu M, Kawauchi J, Sunamori M, Marumo F, Kitajima S, Ikeda MA. Tamamori-Adachi M, et al. Circ Res. 2003 Jan 10;92(1):e12-9. doi: 10.1161/01.res.0000049105.15329.1c. Circ Res. 2003. PMID: 12522130 - Activation of cyclin-dependent kinase 4 (cdk4) by mouse MO15-associated kinase.
Matsuoka M, Kato JY, Fisher RP, Morgan DO, Sherr CJ. Matsuoka M, et al. Mol Cell Biol. 1994 Nov;14(11):7265-75. doi: 10.1128/mcb.14.11.7265-7275.1994. Mol Cell Biol. 1994. PMID: 7935441 Free PMC article. - Rb inactivation in cell cycle and cancer: the puzzle of highly regulated activating phosphorylation of CDK4 versus constitutively active CDK-activating kinase.
Paternot S, Bockstaele L, Bisteau X, Kooken H, Coulonval K, Roger PP. Paternot S, et al. Cell Cycle. 2010 Feb 15;9(4):689-99. doi: 10.4161/cc.9.4.10611. Epub 2010 Feb 12. Cell Cycle. 2010. PMID: 20107323 Review. - The cdk-activating kinase (CAK): from yeast to mammals.
Kaldis P. Kaldis P. Cell Mol Life Sci. 1999 Feb;55(2):284-96. doi: 10.1007/s000180050290. Cell Mol Life Sci. 1999. PMID: 10188587 Free PMC article. Review.
Cited by
- Molecular analysis of cyclin D1 modulators PRKN and FBX4 as candidate tumor suppressors in sporadic parathyroid adenomas.
Brewer K, Nip I, Bellizzi J, Costa-Guda J, Arnold A. Brewer K, et al. Endocr Connect. 2021 Mar;10(3):302-308. doi: 10.1530/EC-21-0055. Endocr Connect. 2021. PMID: 33617468 Free PMC article. - Strategies to generate functionally normal neutrophils to reduce infection and infection-related mortality in cancer chemotherapy.
Abdel-Azim H, Sun W, Wu L. Abdel-Azim H, et al. Pharmacol Ther. 2019 Dec;204:107403. doi: 10.1016/j.pharmthera.2019.107403. Epub 2019 Aug 27. Pharmacol Ther. 2019. PMID: 31470030 Free PMC article. Review. - The DNA deaminase APOBEC3B interacts with the cell-cycle protein CDK4 and disrupts CDK4-mediated nuclear import of Cyclin D1.
McCann JL, Klein MM, Leland EM, Law EK, Brown WL, Salamango DJ, Harris RS. McCann JL, et al. J Biol Chem. 2019 Aug 9;294(32):12099-12111. doi: 10.1074/jbc.RA119.008443. Epub 2019 Jun 19. J Biol Chem. 2019. PMID: 31217276 Free PMC article. - CDK4 protein is degraded by anaphase-promoting complex/cyclosome in mitosis and reaccumulates in early G1 phase to initiate a new cell cycle in HeLa cells.
Chen H, Xu X, Wang G, Zhang B, Wang G, Xin G, Liu J, Jiang Q, Zhang H, Zhang C. Chen H, et al. J Biol Chem. 2017 Jun 16;292(24):10131-10141. doi: 10.1074/jbc.M116.773226. Epub 2017 Apr 26. J Biol Chem. 2017. PMID: 28446612 Free PMC article. - Cyclin D1 in the Liver: Role of Noncanonical Signaling in Liver Steatosis and Hormone Regulation.
Núñez KG, Gonzalez-Rosario J, Thevenot PT, Cohen AJ. Núñez KG, et al. Ochsner J. 2017 Spring;17(1):56-65. Ochsner J. 2017. PMID: 28331449 Free PMC article.
References
- Cell. 1992 Sep 18;70(6):993-1006 - PubMed
- Cell. 1992 Oct 30;71(3):505-14 - PubMed
- Semin Cell Biol. 1991 Aug;2(4):261-70 - PubMed
- Cell. 1993 May 7;73(3):487-97 - PubMed
- Genes Dev. 1993 May;7(5):812-21 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials