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. 1997 Jun;17(6):3408–3417. doi: 10.1128/mcb.17.6.3408

Plk is a functional homolog of Saccharomyces cerevisiae Cdc5, and elevated Plk activity induces multiple septation structures.

K S Lee 1, R L Erikson 1
PMCID: PMC232194  PMID: 9154840

Abstract

Plk is a mammalian serine/threonine protein kinase whose activity peaks at the onset of M phase. It is closely related to other mammalian kinases, Snk, Fnk, and Prk, as well as to Xenopus laevis Plx1, Drosophila melanogaster polo, Schizosaccharomyces pombe Plo1, and Saccharomyces cerevisiae Cdc5. The M phase of the cell cycle is a highly coordinated process which insures the equipartition of genetic and cellular materials during cell division. To enable understanding of the function of Plk during M phase progression, various Plk mutants were generated and expressed in Sf9 cells and budding yeast. In vitro kinase assays with Plk immunoprecipitates prepared from Sf9 cells indicate that Glu206 and Thr210 play equally important roles for Plk activity and that replacement of Thr210 with a negatively charged residue elevates Plk specific activity. Ectopic expression of wild-type Plk (Plk WT) complements the cell division defect associated with the cdc5-1 mutation in S. cerevisiae. The degree of complementation correlates closely with the Plk activity measured in vitro, as it is enhanced by a mutationally activated Plk, T210D, but is not observed with the inactive forms K82M, D194N, and D194R. In a CDC5 wild-type background, expression of Plk WT or T210D, but not of inactive forms, induced a sharp accumulation of cells in G1. Consistent with elevated Plk activity, this phenomenon was enhanced by the C-terminally deleted forms WT deltaC and T210D deltaC. Expression of T210D also induced a class of cells with unusually elongated buds which developed multiple septal structures. This was not observed with the C-terminally deleted form T210D deltaC, however. It appears that the C terminus of Plk is not required for the observed cell cycle influence but may be important for polarized cell growth and septal structure formation.

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Selected References

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  1. Beach D., Durkacz B., Nurse P. Functionally homologous cell cycle control genes in budding and fission yeast. Nature. 1982 Dec 23;300(5894):706–709. doi: 10.1038/300706a0. [DOI] [PubMed] [Google Scholar]
  2. Chant J. Septin scaffolds and cleavage planes in Saccharomyces. Cell. 1996 Jan 26;84(2):187–190. doi: 10.1016/s0092-8674(00)80972-1. [DOI] [PubMed] [Google Scholar]
  3. Clay F. J., McEwen S. J., Bertoncello I., Wilks A. F., Dunn A. R. Identification and cloning of a protein kinase-encoding mouse gene, Plk, related to the polo gene of Drosophila. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):4882–4886. doi: 10.1073/pnas.90.11.4882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crews C. M., Alessandrini A., Erikson R. L. The primary structure of MEK, a protein kinase that phosphorylates the ERK gene product. Science. 1992 Oct 16;258(5081):478–480. doi: 10.1126/science.1411546. [DOI] [PubMed] [Google Scholar]
  5. Donohue P. J., Alberts G. F., Guo Y., Winkles J. A. Identification by targeted differential display of an immediate early gene encoding a putative serine/threonine kinase. J Biol Chem. 1995 Apr 28;270(17):10351–10357. doi: 10.1074/jbc.270.17.10351. [DOI] [PubMed] [Google Scholar]
  6. Drubin D. G., Nelson W. J. Origins of cell polarity. Cell. 1996 Feb 9;84(3):335–344. doi: 10.1016/s0092-8674(00)81278-7. [DOI] [PubMed] [Google Scholar]
  7. Fenton B., Glover D. M. A conserved mitotic kinase active at late anaphase-telophase in syncytial Drosophila embryos. Nature. 1993 Jun 17;363(6430):637–640. doi: 10.1038/363637a0. [DOI] [PubMed] [Google Scholar]
  8. Fode C., Motro B., Yousefi S., Heffernan M., Dennis J. W. Sak, a murine protein-serine/threonine kinase that is related to the Drosophila polo kinase and involved in cell proliferation. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6388–6392. doi: 10.1073/pnas.91.14.6388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  10. Golsteyn R. M., Schultz S. J., Bartek J., Ziemiecki A., Ried T., Nigg E. A. Cell cycle analysis and chromosomal localization of human Plk1, a putative homologue of the mitotic kinases Drosophila polo and Saccharomyces cerevisiae Cdc5. J Cell Sci. 1994 Jun;107(Pt 6):1509–1517. doi: 10.1242/jcs.107.6.1509. [DOI] [PubMed] [Google Scholar]
  11. Haarer B. K., Pringle J. R. Immunofluorescence localization of the Saccharomyces cerevisiae CDC12 gene product to the vicinity of the 10-nm filaments in the mother-bud neck. Mol Cell Biol. 1987 Oct;7(10):3678–3687. doi: 10.1128/mcb.7.10.3678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hamanaka R., Maloid S., Smith M. R., O'Connell C. D., Longo D. L., Ferris D. K. Cloning and characterization of human and murine homologues of the Drosophila polo serine-threonine kinase. Cell Growth Differ. 1994 Mar;5(3):249–257. [PubMed] [Google Scholar]
  13. Hamanaka R., Smith M. R., O'Connor P. M., Maloid S., Mihalic K., Spivak J. L., Longo D. L., Ferris D. K. Polo-like kinase is a cell cycle-regulated kinase activated during mitosis. J Biol Chem. 1995 Sep 8;270(36):21086–21091. doi: 10.1074/jbc.270.36.21086. [DOI] [PubMed] [Google Scholar]
  14. Hardy C. F., Pautz A. A novel role for Cdc5p in DNA replication. Mol Cell Biol. 1996 Dec;16(12):6775–6782. doi: 10.1128/mcb.16.12.6775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Holtrich U., Wolf G., Bräuninger A., Karn T., Böhme B., Rübsamen-Waigmann H., Strebhardt K. Induction and down-regulation of PLK, a human serine/threonine kinase expressed in proliferating cells and tumors. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1736–1740. doi: 10.1073/pnas.91.5.1736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Huang W., Erikson R. L. Constitutive activation of Mek1 by mutation of serine phosphorylation sites. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8960–8963. doi: 10.1073/pnas.91.19.8960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Johnston G. C., Pringle J. R., Hartwell L. H. Coordination of growth with cell division in the yeast Saccharomyces cerevisiae. Exp Cell Res. 1977 Mar 1;105(1):79–98. doi: 10.1016/0014-4827(77)90154-9. [DOI] [PubMed] [Google Scholar]
  19. Kim H. B., Haarer B. K., Pringle J. R. Cellular morphogenesis in the Saccharomyces cerevisiae cell cycle: localization of the CDC3 gene product and the timing of events at the budding site. J Cell Biol. 1991 Feb;112(4):535–544. doi: 10.1083/jcb.112.4.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kitada K., Johnson A. L., Johnston L. H., Sugino A. A multicopy suppressor gene of the Saccharomyces cerevisiae G1 cell cycle mutant gene dbf4 encodes a protein kinase and is identified as CDC5. Mol Cell Biol. 1993 Jul;13(7):4445–4457. doi: 10.1128/mcb.13.7.4445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kumagai A., Dunphy W. G. Purification and molecular cloning of Plx1, a Cdc25-regulatory kinase from Xenopus egg extracts. Science. 1996 Sep 6;273(5280):1377–1380. doi: 10.1126/science.273.5280.1377. [DOI] [PubMed] [Google Scholar]
  22. Lake R. J., Jelinek W. R. Cell cycle- and terminal differentiation-associated regulation of the mouse mRNA encoding a conserved mitotic protein kinase. Mol Cell Biol. 1993 Dec;13(12):7793–7801. doi: 10.1128/mcb.13.12.7793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lee K. S., Yuan Y. L., Kuriyama R., Erikson R. L. Plk is an M-phase-specific protein kinase and interacts with a kinesin-like protein, CHO1/MKLP-1. Mol Cell Biol. 1995 Dec;15(12):7143–7151. doi: 10.1128/mcb.15.12.7143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Li B., Ouyang B., Pan H., Reissmann P. T., Slamon D. J., Arceci R., Lu L., Dai W. Prk, a cytokine-inducible human protein serine/threonine kinase whose expression appears to be down-regulated in lung carcinomas. J Biol Chem. 1996 Aug 9;271(32):19402–19408. doi: 10.1074/jbc.271.32.19402. [DOI] [PubMed] [Google Scholar]
  25. Llamazares S., Moreira A., Tavares A., Girdham C., Spruce B. A., Gonzalez C., Karess R. E., Glover D. M., Sunkel C. E. polo encodes a protein kinase homolog required for mitosis in Drosophila. Genes Dev. 1991 Dec;5(12A):2153–2165. doi: 10.1101/gad.5.12a.2153. [DOI] [PubMed] [Google Scholar]
  26. Marshall C. J. Signal transduction. Hot lips and phosphorylation of protein kinases. Nature. 1994 Feb 24;367(6465):686–686. doi: 10.1038/367686a0. [DOI] [PubMed] [Google Scholar]
  27. Mendenhall M. D., Richardson H. E., Reed S. I. Dominant negative protein kinase mutations that confer a G1 arrest phenotype. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4426–4430. doi: 10.1073/pnas.85.12.4426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ohkura H., Hagan I. M., Glover D. M. The conserved Schizosaccharomyces pombe kinase plo1, required to form a bipolar spindle, the actin ring, and septum, can drive septum formation in G1 and G2 cells. Genes Dev. 1995 May 1;9(9):1059–1073. doi: 10.1101/gad.9.9.1059. [DOI] [PubMed] [Google Scholar]
  29. Piggott J. R., Rai R., Carter B. L. A bifunctional gene product involved in two phases of the yeast cell cycle. Nature. 1982 Jul 22;298(5872):391–393. doi: 10.1038/298391a0. [DOI] [PubMed] [Google Scholar]
  30. Pringle J. R. Staining of bud scars and other cell wall chitin with calcofluor. Methods Enzymol. 1991;194:732–735. doi: 10.1016/0076-6879(91)94055-h. [DOI] [PubMed] [Google Scholar]
  31. Roberts R. L., Bowers B., Slater M. L., Cabib E. Chitin synthesis and localization in cell division cycle mutants of Saccharomyces cerevisiae. Mol Cell Biol. 1983 May;3(5):922–930. doi: 10.1128/mcb.3.5.922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Russell P., Moreno S., Reed S. I. Conservation of mitotic controls in fission and budding yeasts. Cell. 1989 Apr 21;57(2):295–303. doi: 10.1016/0092-8674(89)90967-7. [DOI] [PubMed] [Google Scholar]
  33. Siliciano P. G., Tatchell K. Transcription and regulatory signals at the mating type locus in yeast. Cell. 1984 Jul;37(3):969–978. doi: 10.1016/0092-8674(84)90431-8. [DOI] [PubMed] [Google Scholar]
  34. Simmons D. L., Neel B. G., Stevens R., Evett G., Erikson R. L. Identification of an early-growth-response gene encoding a novel putative protein kinase. Mol Cell Biol. 1992 Sep;12(9):4164–4169. doi: 10.1128/mcb.12.9.4164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Slater M. L., Bowers B., Cabib E. Formation of septum-like structures at locations remote from the budding sites in cytokinesis-defective mutants of Saccharomyces cerevisiae. J Bacteriol. 1985 May;162(2):763–767. doi: 10.1128/jb.162.2.763-767.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sunkel C. E., Glover D. M. polo, a mitotic mutant of Drosophila displaying abnormal spindle poles. J Cell Sci. 1988 Jan;89(Pt 1):25–38. doi: 10.1242/jcs.89.1.25. [DOI] [PubMed] [Google Scholar]
  37. Surana U., Robitsch H., Price C., Schuster T., Fitch I., Futcher A. B., Nasmyth K. The role of CDC28 and cyclins during mitosis in the budding yeast S. cerevisiae. Cell. 1991 Apr 5;65(1):145–161. doi: 10.1016/0092-8674(91)90416-v. [DOI] [PubMed] [Google Scholar]
  38. Taylor S. S., Knighton D. R., Zheng J., Sowadski J. M., Gibbs C. S., Zoller M. J. A template for the protein kinase family. Trends Biochem Sci. 1993 Mar;18(3):84–89. doi: 10.1016/0968-0004(93)80001-r. [DOI] [PubMed] [Google Scholar]
  39. 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]

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