Russo, G. L., Vandenberg, M. T., Yu, I. J., Bae, Y. S., Franza, B. R., Marshak, D. R. (October 1992) Casein Kinase-Ii Phosphorylates P34(Cdc2) Kinase in G1 Phase of the Hela-Cell Division Cycle. Journal of Biological Chemistry, 267 (28). pp. 20317-20325. ISSN 0021-9258
Abstract
The activity of p34cdc2 kinase is regulated in the phases of vertebrate cell cycle by mechanisms of phosphorylation and dephosphorylation. In this paper, we demonstrate that casein kinase II (CKII) phosphorylates p34cdc2 in vivo and in vitro at Ser 39 during the G1 phase of HeLa cell division cycle. Human p34cdc2 shows a typical phosphorylation sequence motif site for CKII at Ser39 (ES39EEE). In our experiments, either p34cdc2 expressed and purified from bacteria or p34cdc2 immunoprecipitated from HeLa cells enriched in G1 by elutriation were substrates for in vitro phosphorylation by CKII. Phosphoamino acid analysis, N-chlorosuccinimide mapping, and two-dimensional tryptic mapping of p34cdc2 phosphorylated in vitro were performed to determine the phosphorylation site. A synthetic peptide spanning residues 33-50 of human p34cdc2, including the CKII site, was used to map the site. In addition, phosphorylation at Ser39 also occurs in vivo, since p34cdc2 is phosphorylated during G1 on serine, and its two-dimensional tryptic map shows two phosphopeptides that comigrate exactly with the synthetic peptides used as standard.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | P34CDC2 PROTEIN-KINASE MITOSIS-SPECIFIC PHOSPHORYLATION RAT-LIVER CYTOSOL SV40 T-ANTIGEN FISSION YEAST PROMOTING FACTOR SCHIZOSACCHAROMYCES-POMBE TYROSINE PHOSPHORYLATION SACCHAROMYCES-CEREVISIAE RETINOBLASTOMA PROTEIN |
| Subjects: | organs, tissues, organelles, cell types and functions > cell types and functions > cell functions > cell cycle bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > kinase bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein expression > phosphorylation |
| CSHL Authors: | |
| Communities: | CSHL labs |
| Depositing User: | Matt Covey |
| Date: | October 1992 |
| Date Deposited: | 28 Sep 2015 19:11 |
| Last Modified: | 28 Sep 2015 19:11 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/31813 |
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