Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1997 Mar;113(3):841–852. doi: 10.1104/pp.113.3.841

The cdc2Ms Kinase Is Differently Regulated in the Cytoplasm and in the Nucleus.

L Bogre 1, K Zwerger 1, I Meskiene 1, P Binarova 1, V Csizmadia 1, C Planck 1, E Wagner 1, H Hirt 1, E Heberle-Bors 1
PMCID: PMC158204  PMID: 12223648

Abstract

To study a cyclin-dependent kinase (CDK) from alfalfa (Medicago sativa L.), an antibody was raised against the C-terminal 16 amino acids of the protein cdc2aMs. The cdc2Ms protein was immunopurified with this antibody and its histone kinase activity was measured. The cdc2Ms kinase is activated at the G1/S transition when phosphate-starved cells from the G0 phase re-enter the cell cycle and remain active as cells transit the S, G2, and M phases, indicating that the same CDK regulates all of these phases in alfalfa. In contrast, when cdc2Ms kinase was purified by binding to p13suc1, it was active only in the G2 and M phases. In immunoblots the C-terminal antibody detected an equal amount of the cdc2Ms protein in the cytoplasm and in the nucleus. By indirect immunofluorescence, however, the cytoplasmic form of cdc2Ms could not be found in the S phase of the cells, indicating that the epitope for the cdc2 antibody is not accessible. Binding of putative inhibitor proteins to cdc2 was shown by inactivation of purified plant CDK when cell extracts were added. Furthermore, purified CDK inhibitors, such as the mouse p27kip1 and the yeast p40sic1, blocked the purified plant CDK activity.

Full Text

The Full Text of this article is available as a PDF (4.5 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bakó L., Nuotio S., Dudits D., Schell J., Koncz C. RNAPII: a specific target for the cell cycle kinase complex. Results Probl Cell Differ. 1994;20:25–64. doi: 10.1007/978-3-540-48037-2_2. [DOI] [PubMed] [Google Scholar]
  2. Bögre L., Jonak C., Mink M., Meskiene I., Traas J., Ha D. T., Swoboda I., Plank C., Wagner E., Heberle-Bors E. Developmental and cell cycle regulation of alfalfa nucMs1, a plant homolog of the yeast Nsr1 and mammalian nucleolin. Plant Cell. 1996 Mar;8(3):417–428. [PMC free article] [PubMed] [Google Scholar]
  3. Colasanti J., Cho S. O., Wick S., Sundaresan V. Localization of the Functional p34cdc2 Homolog of Maize in Root Tip and Stomatal Complex Cells: Association with Predicted Division Sites. Plant Cell. 1993 Sep;5(9):1101–1111. doi: 10.1105/tpc.5.9.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Colasanti J., Tyers M., Sundaresan V. Isolation and characterization of cDNA clones encoding a functional p34cdc2 homologue from Zea mays. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3377–3381. doi: 10.1073/pnas.88.8.3377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feiler H. S., Jacobs T. W. Cell division in higher plants: a cdc2 gene, its 34-kDa product, and histone H1 kinase activity in pea. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5397–5401. doi: 10.1073/pnas.87.14.5397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ferreira P. C., Hemerly A. S., Villarroel R., Van Montagu M., Inzé D. The Arabidopsis functional homolog of the p34cdc2 protein kinase. Plant Cell. 1991 May;3(5):531–540. doi: 10.1105/tpc.3.5.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fobert P. R., Gaudin V., Lunness P., Coen E. S., Doonan J. H. Distinct classes of cdc2-related genes are differentially expressed during the cell division cycle in plants. Plant Cell. 1996 Sep;8(9):1465–1476. doi: 10.1105/tpc.8.9.1465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grafi G., Larkins B. A. Endoreduplication in maize endosperm: involvement of m phase--promoting factor inhibition and induction of s phase--related kinases. Science. 1995 Sep 1;269(5228):1262–1264. doi: 10.1126/science.269.5228.1262. [DOI] [PubMed] [Google Scholar]
  9. Hashimoto J., Hirabayashi T., Hayano Y., Hata S., Ohashi Y., Suzuka I., Utsugi T., Toh-e A., Kikuchi Y. Isolation and characterization of cDNA clones encoding cdc2 homologues from Oryza sativa: a functional homologue and cognate variants. Mol Gen Genet. 1992 May;233(1-2):10–16. doi: 10.1007/BF00587555. [DOI] [PubMed] [Google Scholar]
  10. Hata S. cDNA cloning of a novel cdc2+/CDC28-related protein kinase from rice. FEBS Lett. 1991 Feb 11;279(1):149–152. doi: 10.1016/0014-5793(91)80271-4. [DOI] [PubMed] [Google Scholar]
  11. Hemerly A. S., Ferreira P., de Almeida Engler J., Van Montagu M., Engler G., Inzé D. cdc2a expression in Arabidopsis is linked with competence for cell division. Plant Cell. 1993 Dec;5(12):1711–1723. doi: 10.1105/tpc.5.12.1711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hemerly A., Engler J. de A., Bergounioux C., Van Montagu M., Engler G., Inzé D., Ferreira P. Dominant negative mutants of the Cdc2 kinase uncouple cell division from iterative plant development. EMBO J. 1995 Aug 15;14(16):3925–3936. doi: 10.1002/j.1460-2075.1995.tb00064.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hirayama T., Imajuku Y., Anai T., Matsui M., Oka A. Identification of two cell-cycle-controlling cdc2 gene homologs in Arabidopsis thaliana. Gene. 1991 Sep 15;105(2):159–165. doi: 10.1016/0378-1119(91)90146-3. [DOI] [PubMed] [Google Scholar]
  14. Hirt H., Páy A., Bögre L., Meskiene I., Heberle-Bors E. cdc2MsB, a cognate cdc2 gene from alfalfa, complements the G1/S but not the G2/M transition of budding yeast cdc28 mutants. Plant J. 1993 Jul;4(1):61–69. doi: 10.1046/j.1365-313x.1993.04010061.x. [DOI] [PubMed] [Google Scholar]
  15. Hirt H., Páy A., Györgyey J., Bakó L., Németh K., Bögre L., Schweyen R. J., Heberle-Bors E., Dudits D. Complementation of a yeast cell cycle mutant by an alfalfa cDNA encoding a protein kinase homologous to p34cdc2. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1636–1640. doi: 10.1073/pnas.88.5.1636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Imajuku Y., Hirayama T., Endoh H., Oka A. Exon-intron organization of the Arabidopsis thaliana protein kinase genes CDC2a and CDC2b. FEBS Lett. 1992 Jun 8;304(1):73–77. doi: 10.1016/0014-5793(92)80592-5. [DOI] [PubMed] [Google Scholar]
  17. John P. C., Sek F. J., Lee M. G. A homolog of the cell cycle control protein p34cdc2 participates in the division cycle of Chlamydomonas, and a similar protein is detectable in higher plants and remote taxa. Plant Cell. 1989 Dec;1(12):1185–1193. doi: 10.1105/tpc.1.12.1185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kaffman A., Herskowitz I., Tjian R., O'Shea E. K. Phosphorylation of the transcription factor PHO4 by a cyclin-CDK complex, PHO80-PHO85. Science. 1994 Feb 25;263(5150):1153–1156. doi: 10.1126/science.8108735. [DOI] [PubMed] [Google Scholar]
  19. Martinez M. C., Jørgensen J. E., Lawton M. A., Lamb C. J., Doerner P. W. Spatial pattern of cdc2 expression in relation to meristem activity and cell proliferation during plant development. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7360–7364. doi: 10.1073/pnas.89.16.7360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Meijer L., Arion D., Golsteyn R., Pines J., Brizuela L., Hunt T., Beach D. Cyclin is a component of the sea urchin egg M-phase specific histone H1 kinase. EMBO J. 1989 Aug;8(8):2275–2282. doi: 10.1002/j.1460-2075.1989.tb08353.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mendenhall M. D. An inhibitor of p34CDC28 protein kinase activity from Saccharomyces cerevisiae. Science. 1993 Jan 8;259(5092):216–219. doi: 10.1126/science.8421781. [DOI] [PubMed] [Google Scholar]
  22. Meskiene I., Bögre L., Dahl M., Pirck M., Ha D. T., Swoboda I., Heberle-Bors E., Ammerer G., Hirt H. cycMs3, a novel B-type alfalfa cyclin gene, is induced in the G0-to-G1 transition of the cell cycle. Plant Cell. 1995 Jun;7(6):759–771. doi: 10.1105/tpc.7.6.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Meyerson M., Enders G. H., Wu C. L., Su L. K., Gorka C., Nelson C., Harlow E., Tsai L. H. A family of human cdc2-related protein kinases. EMBO J. 1992 Aug;11(8):2909–2917. doi: 10.1002/j.1460-2075.1992.tb05360.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Miao G. H., Hong Z., Verma D. P. Two functional soybean genes encoding p34cdc2 protein kinases are regulated by different plant developmental pathways. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):943–947. doi: 10.1073/pnas.90.3.943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Morgan D. O. Principles of CDK regulation. Nature. 1995 Mar 9;374(6518):131–134. doi: 10.1038/374131a0. [DOI] [PubMed] [Google Scholar]
  26. Perennes C., Qin L. X., Glab N., Bergounioux C. Petunia p34cdc2 protein kinase activity in G2/M cells obtained with a reversible cell cycle inhibitor, mimosine. FEBS Lett. 1993 Oct 25;333(1-2):141–145. doi: 10.1016/0014-5793(93)80392-8. [DOI] [PubMed] [Google Scholar]
  27. Polyak K., Lee M. H., Erdjument-Bromage H., Koff A., Roberts J. M., Tempst P., Massagué J. Cloning of p27Kip1, a cyclin-dependent kinase inhibitor and a potential mediator of extracellular antimitogenic signals. Cell. 1994 Jul 15;78(1):59–66. doi: 10.1016/0092-8674(94)90572-x. [DOI] [PubMed] [Google Scholar]
  28. Poon R. Y., Yamashita K., Adamczewski J. P., Hunt T., Shuttleworth J. The cdc2-related protein p40MO15 is the catalytic subunit of a protein kinase that can activate p33cdk2 and p34cdc2. EMBO J. 1993 Aug;12(8):3123–3132. doi: 10.1002/j.1460-2075.1993.tb05981.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rosenblatt J., Gu Y., Morgan D. O. Human cyclin-dependent kinase 2 is activated during the S and G2 phases of the cell cycle and associates with cyclin A. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2824–2828. doi: 10.1073/pnas.89.7.2824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schwob E., Böhm T., Mendenhall M. D., Nasmyth K. The B-type cyclin kinase inhibitor p40SIC1 controls the G1 to S transition in S. cerevisiae. Cell. 1994 Oct 21;79(2):233–244. doi: 10.1016/0092-8674(94)90193-7. [DOI] [PubMed] [Google Scholar]
  31. Sherr C. J., Roberts J. M. Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev. 1995 May 15;9(10):1149–1163. doi: 10.1101/gad.9.10.1149. [DOI] [PubMed] [Google Scholar]
  32. Stern B., Ried G., Clegg N. J., Grigliatti T. A., Lehner C. F. Genetic analysis of the Drosophila cdc2 homolog. Development. 1993 Jan;117(1):219–232. doi: 10.1242/dev.117.1.219. [DOI] [PubMed] [Google Scholar]
  33. van den Heuvel S., Harlow E. Distinct roles for cyclin-dependent kinases in cell cycle control. Science. 1993 Dec 24;262(5142):2050–2054. doi: 10.1126/science.8266103. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES