Mol. Cell. Biol., Jan 1995, 345-350, Vol 15, No. 1
Copyright © 1995, American Society for Microbiology

Effects of phosphorylation by CAK on cyclin binding by CDC2 and CDK2

D Desai, HC Wessling, RP Fisher and DO Morgan
Department of Physiology, University of California, San Francisco 94143- 0444.

The cyclin-dependent protein kinases (CDKs) are activated by association with cyclins and by phosphorylation at a conserved threonine residue by the CDK-activating kinase (CAK). We have studied the binding of various human CDK and cyclin subunits in vitro, using purified proteins derived from baculovirus-infected insect cells. We find that most CDK-cyclin complexes known to exist in human cells (CDC2- cyclin B, CDK2-cyclin A, and CDK2-cyclin E) form with high affinity in the absence of phosphorylation or other cellular components. One complex (CDC2-cyclin A) forms with high affinity only after CAK- mediated phosphorylation of CDC2 at the activating threonine residue. CDC2 does not bind with high affinity to cyclin E in vitro, even after phosphorylation of the CDC2 subunit. Thus, phosphorylation is of varying importance in the formation of high-affinity CDK-cyclin complexes.

This article has been cited by other articles:

• Fisher, R. P. (2005). Secrets of a double agent: CDK7 in cell-cycle control and transcription. J. Cell Sci. 118: 5171-5180 [Abstract] [Full Text]  
• Katayama, K., Fujita, N., Tsuruo, T. (2005). Akt/Protein Kinase B-Dependent Phosphorylation and Inactivation of WEE1Hu Promote Cell Cycle Progression at G2/M Transition. Mol. Cell. Biol. 25: 5725-5737 [Abstract] [Full Text]  
• Gondeau, C., Gerbal-Chaloin, S., Bello, P., Aldrian-Herrada, G., Morris, M. C., Divita, G. (2005). Design of a Novel Class of Peptide Inhibitors of Cyclin-dependent Kinase/Cyclin Activation. J. Biol. Chem. 280: 13793-13800 [Abstract] [Full Text]  
• Major, M. L., Lepe, R., Costa, R. H. (2004). Forkhead Box M1B Transcriptional Activity Requires Binding of Cdk-Cyclin Complexes for Phosphorylation-Dependent Recruitment of p300/CBP Coactivators. Mol. Cell. Biol. 24: 2649-2661 [Abstract] [Full Text]  
• Ohlmeyer, J. T., Schupbach, T. (2003). Encore facilitates SCF-Ubiquitin-proteasome-dependent proteolysis during Drosophila oogenesis. Development 130: 6339-6349 [Abstract] [Full Text]  
• Prokhorova, T. A., Mowrer, K., Gilbert, C. H., Walter, J. C. (2003). DNA replication of mitotic chromatin in Xenopus egg extracts. Proc. Natl. Acad. Sci. USA 100: 13241-13246 [Abstract] [Full Text]  
• Santaguida, M., Ding, Q., Berube, G., Truscott, M., Whyte, P., Nepveu, A. (2001). Phosphorylation of the CCAAT Displacement Protein (CDP)/Cux Transcription Factor by Cyclin A-Cdk1 Modulates Its DNA Binding Activity in G2. J. Biol. Chem. 276: 45780-45790 [Abstract] [Full Text]  
• CLEMENT, A., HENRION-CAUDE, A., BESNARD, V., CORROYER, S. (2001). Role of Cyclins in Epithelial Response to Oxidants. Am. J. Respir. Crit. Care Med. 164: S81-84 [Abstract] [Full Text]  
• Matt Kim, J., McGaughy, J. T., Kent Bogle, R., Ravnik, S. E. (2001). Meiotic Expression of the Cyclin H/Cdk7 Complex in Male Germ Cells of the Mouse. Biol. Reprod. 64: 1400-1408 [Abstract] [Full Text]  
• Janss, A. J., Maity, A., Tang, C.-B., Muschel, R. J., McKenna, W. G., Sutton, L., Phillips, P. C. (2001). Decreased cyclin B1 expression contributes to G2 delay in human brain tumor cells after treatment with camptothecin. Neuro Oncol 3: 11-21 [Abstract]  
• Bond, G. L., Prives, C., Manley, J. L. (2000). Poly(A) Polymerase Phosphorylation Is Dependent on Novel Interactions with Cyclins. Mol. Cell. Biol. 20: 5310-5320 [Abstract] [Full Text]  
• Ross, K. E., Kaldis, P., Solomon, M. J. (2000). Activating Phosphorylation of the Saccharomyces cerevisiae Cyclin-dependent Kinase, Cdc28p, Precedes Cyclin Binding. Mol. Biol. Cell 11: 1597-1609 [Abstract] [Full Text]  
• Farrell, A., Morgan, D. O. (2000). Cdc37 Promotes the Stability of Protein Kinases Cdc28 and Cak1. Mol. Cell. Biol. 20: 749-754 [Abstract] [Full Text]  
• Kohn, K. W. (1999). Molecular Interaction Map of the Mammalian Cell Cycle Control and DNA Repair Systems. Mol. Biol. Cell 10: 2703-2734 [Abstract] [Full Text]  
• Kimmelman, J., Kaldis, P., Hengartner, C. J., Laff, G. M., Koh, S. S., Young, R. A., Solomon, M. J. (1999). Activating Phosphorylation of the Kin28p Subunit of Yeast TFIIH by Cak1p. Mol. Cell. Biol. 19: 4774-4787 [Abstract] [Full Text]  
• Cross, F. R., Levine, K. (1998). Molecular Evolution Allows Bypass of the Requirement for Activation Loop Phosphorylation of the Cdc28 Cyclin-Dependent Kinase. Mol. Cell. Biol. 18: 2923-2931 [Abstract] [Full Text]  
• Harper, J. W., Elledge, S. J. (1998). The role of Cdk7 in CAK function, a retro-retrospective. Genes Dev. 12: 285-289 [Full Text]  
• Larochelle, S., Pandur, J., Fisher, R. P., Salz, H. K., Suter, B. (1998). Cdk7 is essential for mitosis and for in vivo Cdk-activating kinase activity. Genes Dev. 12: 370-381 [Abstract] [Full Text]  
• Hawley, S. A., Selbert, M. A., Goldstein, E. G., Edelman, A. M., Carling, D., Hardie, D. G. (1995). 5`-AMP Activates the AMP-activated Protein Kinase Cascade, and Ca[IMAGE]/Calmodulin Activates the Calmodulin-dependent Protein Kinase I Cascade, via Three Independent Mechanisms. J. Biol. Chem. 270: 27186-27191 [Abstract] [Full Text]  
• Kaldis, P., Cheng, A., Solomon, M. J. (2000). The Effects of Changing the Site of Activating Phosphorylation in CDK2 from Threonine to Serine. J. Biol. Chem. 275: 32578-32584 [Abstract] [Full Text]  
• Tokuyama, Y., Horn, H. F., Kawamura, K., Tarapore, P., Fukasawa, K. (2001). Specific Phosphorylation of Nucleophosmin on Thr199 by Cyclin- dependent Kinase 2-Cyclin E and Its Role in Centrosome Duplication. J. Biol. Chem. 276: 21529-21537 [Abstract] [Full Text]  
• Derjuga, A., Richard, C., Crosato, M., Wright, P. S., Chalifour, L., Valdez, J., Barraso, A., Crissman, H. A., Nishioka, W., Bradbury, E. M., Th'ng, J. P. H. (2001). Expression of p21Waf1/Cip1 and Cyclin D1 Is Increased in Butyrate-resistant HeLa Cells. J. Biol. Chem. 276: 37815-37820 [Abstract] [Full Text]