Activation of cyclin-dependent kinase 2 by full length and low molecular weight forms of cyclin E in breast cancer cells.
Cyclin E, a positive regulator of the cell cycle, controls the transition of cells from G(1) to S phase. Deregulation of the G(1)-S checkpoint contributes to uncontrolled cell division, a hallmark of cancer. We have reported previously that cyclin E is overexpressed in breast cancer and such overexpression is usually accompanied by the appearance of low molecular weight isoforms of cyclin E protein, which are not present in normal cells. Furthermore, we have shown that the expression of cyclin E low molecular weight isoforms can be used as a reliable prognostic marker for breast cancer to predict patient outcome. In this study we examined the role of cyclin E in directly activating cyclin-dependent kinase (CDK) 2. For this purpose, a series of N-terminal deleted forms of cyclin E corresponding to the low molecular weight forms detected only in cancer cells were translated in vitro and mixed with cell extracts. These tumor-specific N-terminal deleted forms of cyclin E are able to activate CDK2. Addition of cyclin E into both normal and tumor cell extracts was shown to increase the levels of CDK2 activity, along with an increase in the amount of phosphorylated CDK2. The increase in CDK2 activity was because of cyclin E binding to endogenous CDK2 in complex with endogenous cyclin E, cyclin A, or unbound CDK2. The increase in CDK2 phosphorylation was through a pathway involving cyclin-activating kinase, but addition of cyclin E to an extract containing unphosphorylated CDK2 can still lead to increase in CDK2 activity. Our data suggest that the ability of high levels of full-length and low molecular weight forms of cyclin E to activate CDK2 may be one mechanism that leads to the constitutive activation of cyclin E.CDK2 complexes leading to G(1)/S deregulation and tumor progression.
Harwell
Richard M
RM
Division of Molecular Medicine, Wadsworth Center, SUNY Albany, Albany, NY 12201-0509, USA.
Mull
Benjamin B
BB
Porter
Donald C
DC
Keyomarsi
Khandan
K
D000818:Animals; D014408:Biomarkers, Tumor; D001943:Breast Neoplasms / Q000378:metabolism*; D042846:CDC2-CDC28 Kinases / Q000378:metabolism*; D002455:Cell Division; D002460:Cell Line; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D019927:Cyclin E / Q000737:chemistry* / Q000378:metabolism*; D051357:Cyclin-Dependent Kinase 2; D018450:Disease Progression; D004789:Enzyme Activation; D016193:G1 Phase; D006801:Humans; D007313:Insecta; D008954:Models, Biological; D009369:Neoplasms / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D011485:Protein Binding; D014176:Protein Biosynthesis; D020033:Protein Isoforms; D011494:Protein Kinases / Q000378:metabolism; D011817:Rabbits; D016196:S Phase; D014158:Transcription, Genetic
D014408:Biomarkers, Tumor; D019927:Cyclin E; D020033:Protein Isoforms; D011494:Protein Kinases; C102333:histone H1 kinase; D042846:CDC2-CDC28 Kinases; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2