Journal article
D-type cyclin-dependent kinase activity in mammalian cells
Molecular and cellular biology, Vol.14(3), pp.2066-2076
03/1994
DOI: 10.1128/mcb.14.3.2066-2076.1994
PMCID: PMC358567
PMID: 8114738
Abstract
D-type cyclin-dependent kinase activities have not so far been detected in mammalian cells. Lysis of rodent fibroblasts, mouse macrophages, or myeloid cells with Tween 20 followed by precipitation with antibodies to cyclins D1, D2, and D3 or to their major catalytic partner, cyclin-dependent kinase 4 (cdk4), yielded kinase activities in immune complexes which readily phosphorylated the retinoblastoma protein (pRb) but not histone H1 or casein. Virtually all cyclin D1-dependent kinase activity in proliferating macrophages and fibroblasts could be attributed to cdk4. When quiescent cells were stimulated by growth factors to enter the cell cycle, cyclin D1-dependent kinase activity was first detected in mid G1, reached a maximum near the G1/S transition, and remained elevated in proliferating cells. The rate of appearance of kinase activity during G1 phase lagged significantly behind cyclin induction and correlated with the more delayed accumulation of cdk4 and formation of cyclin D1-cdk4 complexes. Thus, cyclin D1-associated kinase activity was not detected during the G0-to-G1 transition, which occurs within the first few hours following growth factor stimulation. Rodent fibroblasts engineered to constitutively overexpress either cyclin D1 alone or cyclin D3 together with cdk4 exhibited greatly elevated cyclin D-dependent kinase activity, which remained absent in quiescent cells but rose to supraphysiologic levels as cells progressed through G1. Therefore, despite continued enforced overproduction of cyclins and cdk4, the assembly of cyclin D-cdk4 complexes and the appearance of their kinase activities remained dependent upon serum stimulation, indicating that upstream regulators must govern formation of the active enzymes.
Details
- Title: Subtitle
- D-type cyclin-dependent kinase activity in mammalian cells
- Creators
- Hitoshi Matsushime - Department of Genetics, University of Tokyo, JapanDawn E Quelle - Department of Genetics, University of Tokyo, JapanSheila A Shurtleff - Department of Genetics, University of Tokyo, JapanMasabumi Shibuya - Department of Genetics, University of Tokyo, JapanCharles J Sherr - Department of Genetics, University of Tokyo, JapanJun-Ya Kato - Department of Genetics, University of Tokyo, Japan
- Resource Type
- Journal article
- Publication Details
- Molecular and cellular biology, Vol.14(3), pp.2066-2076
- DOI
- 10.1128/mcb.14.3.2066-2076.1994
- PMID
- 8114738
- PMCID
- PMC358567
- ISSN
- 0270-7306
- eISSN
- 1098-5549
- Language
- English
- Date published
- 03/1994
- Academic Unit
- Pathology; Neuroscience and Pharmacology
- Record Identifier
- 9984040589902771
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