Skip to main content
Log in

Themet1 mutation inChlamydomonas reinhardtii causes arrest at mitotic metaphase with persisting p34cdc2-like H1 histone kinase activity that can promote mitosis when injected into higher-plant cells

  • Original Papers
  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

Themet1 mutation inChlamydomonas reinhardtii causes metaphase arrest. Arrested cells have disassembled cortical microtubules, a fully assembled spindle, condensed and aligned metaphase chromosomes and abundant mitotic phosphoproteins recognised by MPM-2 antibody in the nuclear region. Protein purified by affinity for the mitotic protein p13suc1 contains p34cdc2-like H1 histone kinase activity at times when control cells have inactivated this enzyme. The active enzyme, when microinjected intoTradescantia stamen hair cells, accelerated progress through prophase to normal completion of mitosis, indicating that the mutation did not disable the mitotic Cdc2 protein kinase enzyme complex. The mutation prevented the normal lowering of this kinase activity that accompanies anaphase. A defect at time of mitosis rather than earlier in the cycle was indicated by temperature shifting of synchronous cells, which identified the earliest faulty progress as occurring near the beginning of mitosis and the time at which the essential function is completed near the end of mitosis. Themet1 gene mapped approximately 33 cM fromery-2 and extended the known limits of the linkage group XIV.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Beach D, Durkacz B, Nurse P (1982) Functionally homologous cell cycle control genes in budding and fission yeast. Nature 300: 706–709

    Google Scholar 

  • Chang F, Nurse P (1993) Finishing the cell cycle: control of mitosis and cytokinesis in fission yeast. Trends Genet 9: 333–335

    Google Scholar 

  • Colasanti J, Tyers M, Sundaresan V (1991) Isolation and characterisation of cDNA clones encoding a functional p34cdc2 homologue fromZea mays. Proc Natl Acad Sci USA 88: 3377–3381

    Google Scholar 

  • Cross F (1995) Starting the cell cycle: what's the point. Curr Opin Cell Biol 7: 790–797

    Google Scholar 

  • Donnan L, John PCL (1983) Cell cycle control by timer and sizer inChlamydomonas. Nature 340: 630–633

    Google Scholar 

  • —, Carvill EP, Gilliland TJ, John PCL (1985) The cell cycle ofChlamydomonas andChlorella. New Phytol 99: 1–40

    Google Scholar 

  • Donovan JD, Toyn JH, Johnson AL (1994) p40SDB25, a putative CDK inhibitor, has a role in the M/G1 transition inSaccharomyces cerevisiae. Genes Dev 8: 1640–1653

    Google Scholar 

  • Draetta G, Beach D (1988) Activation of cdc2 protein kinase during mitosis in human cells: cell cycle-dependent phosphorylation and subunit rearrangement. Cell 54: 17–26

    Google Scholar 

  • Dutcher SK, Power J, Galloway RE, Porter ME (1991) Reappraisal of the genetic map ofChlamydomonas reinhardtii. J Hered 82: 295–301

    Google Scholar 

  • Enoch T, Carr A, Nurse P (1992) Fission yeast genes involved in coupling mitosis to completion of DNA replication. Genes Dev 6: 2035–2046

    Google Scholar 

  • Ferriera P, Hemerly A, Van Montague M, Inzé D (1994) Control of plant cell proliferation during plant development. Plant Mol Biol 26: 1289–1303

    Google Scholar 

  • Fobert PR, Coen ES, Murphy GJP, Doonan JH (1994) Patterns of cell division revealed by transcriptional regulation of genes during the cell cycle in plants. EMBO J 13: 616–624

    Google Scholar 

  • Gorman DS, Levine RP (1965) Cytochrome F and plastocyanin: their sequence in the photosynthesis transport chain ofChlamydomonas reinhardtii. Proc Natl Acad Sci USA 54: 1665–1669

    Google Scholar 

  • Harper JDI, John PCL (1986) Coordination of division events in theChlamydomonas cell cycle. Protoplasma 131: 118–130

    Google Scholar 

  • —, Rao PN, John PCL (1990) The mitosis-specific monoclonal antibody MPM-2 recognises phosphoproteins associated with the nuclear envelope inChlamydomonas reinhardtii cells. Eur J Cell Biol 51: 272–278

    Google Scholar 

  • —, McCurdy DW, Sanders MA, Salisbury JL, John PCL (1992) Actin dynamics during the cell cycle inChlamydomonas reinhardtii. Cell Motil Cytoskeleton 22: 117–126

    Google Scholar 

  • —, Wu L, Sakuanrungsirikul S, John PCL (1995) Isolation and partial characterisation of conditional cell division cycle mutants inChlamydomonas. Protoplasma 186: 149–162

    Google Scholar 

  • Harris EH (1989) TheChlamydomonas sourcebook: a comprehensive guide to biology and laboratory use. Academic Press, San Diego

    Google Scholar 

  • Hartwell LH (1991) Twenty-five years of cell cycle genetics. Genetics 129: 975–980

    Google Scholar 

  • —, Weinert T (1989) Checkpoints: controls that ensure the order of cell cycle events. Science 246: 629–634

    Google Scholar 

  • Hayles J, Aves S, Nurse P (1986) Suc 1 is an essential gene involved in both the cell cycle and growth in fission yeast. EMBO J 5: 3373–3379

    Google Scholar 

  • Hepler PK (1985) Calcium restriction prolongs metaphase in dividing stamen hair cells ofTradescantia. J Cell Biol 100: 1363–1368

    Google Scholar 

  • —, Sek FJ, John PCL (1994) Nuclear concentration and mitotic dispersion of the essential cell cycle protein, p13suc1, examined in living cells. Proc Natl Acad Sci USA 91: 2176–2180

    Google Scholar 

  • Hindley J, Phear GA, Stein M, Beach D (1987)Sucl + encodes a predicted 13-kilodalton protein that is essential for cell viability and directly involved in the division cycle ofSchizosaccharomyces pombe. Mol Cell Biol 7: 504–511

    Google Scholar 

  • Hirt H, Pay A, Gyorgyey J, Bako L, Newmeth K, Borge L, Schweyen R J, Heberle-Bors E, Dudits D (1991) Complementation of a yeast cell cycle mutant by an alfalfa cDNA encoding a protein kinase homologous to p34cdc2. Proc Natl Acad Sci USA 88: 1636–1640

    Google Scholar 

  • Hollofnway SL, Glotzer M, King RW, Murray AW (1993) Anaphase is initiated by proteolysis rather than by inactivation of maturationpromoting factor. Cell 73: 1393–1402

    Google Scholar 

  • Howell SH, Naliboff JA (1973) Conditional mutants inChlamydomonas reinhardtii blocked in the vegetative cell cycle 1. An analysis of cell cycle block points. J Cell Biol 57: 760–772

    Google Scholar 

  • Hush JM, Wadsworth P, Callaham DA, Hepler PK (1994) Quantification of the microtubule dynamics in living plant cells using fluorescence redistribution after photobleaching. J Cell Sci 107: 775–784

    Google Scholar 

  • —, Wu L, John PCL, Hepler LH, Hepler PK (1996) Plant mitosis promoting factor disassembles the microtubule preprophase band and accelerates prophase progression inTradescantia. Cell Biol Int 20: 275–287

    Google Scholar 

  • John PCL (1987) Control points inChlamydomonas cell cycle. In: Wiessner W, Robinson DG, Starr RC (eds) Algal development: molecular and cellular aspects. Springer, Berlin Heidelberg New York Tokyo, pp 9–16

    Google Scholar 

  • — (1996) The plant cell cycle: conserved and unique features in mitotic control. Prog Cell Cycle Res 2: 59–72

    Google Scholar 

  • —, Sek FJ, Lee MG (1989) A homologue of the cell cycle control protein p34cdc2 participates in the cell division cycle ofChlamydomonas and a similar protein is detectable in higher plants and remote taxa. Plant Cell 1: 1185–1193

    Google Scholar 

  • — —, Carmichael JP, McCurdy DW (1990) p34cdc2 homologue level, cell division, phytohormone responsiveness and cell differentiation in wheat leaves. J Cell Sci 97: 627–630

    Google Scholar 

  • — —, Hayles J (1991) Association of the plant p34cdc2-like protein with p13suc1: implications for control of cell division cycles in plants. Protoplasma 161: 70–74

    Google Scholar 

  • King RW, Deshaies RJ, Peters J-M, Kirschner MW (1996) How proteoiysis drives the cell cycle. Science 274: 1652–1659

    Google Scholar 

  • Kinoshita N, Yamano H, Niwa H, Yoshida T, Yanagida (1993) Negative regulation of mitosis by the fission yeast protein phosphatase ppa2. Genes Dev 7: 1059–1071

    Google Scholar 

  • Kouchi H, Sekine M, Hata S (1995) Distinct classes of mitotic cyclins are differentially expressed in the soybean shoot apex during the cell cycle. Plant Cell 7: 1143–1155

    Google Scholar 

  • Labib K, Moreno S (1996) Rum1: a CDK inhibitor regulating G1 progression in fission yeast. Trends Cell Biol 6: 62–66

    Google Scholar 

  • Lamb JR, Michaud WA, Sikorski RS, Hieter PA (1994) Cdc16p, Cdc23p and Cdc27p form a complex essential for mitosis. EMBO J 13: 4321–4328

    Google Scholar 

  • Lamb NJC, Fernandez A, Watrin A, Labbé J-C, Cavadoré J-C (1990) Microinjection of p34cdc2 kinase induces marked changes in cell shape, cytoskeletal organisation and chromatin structure in mammalian fibroblasts. Cell 60: 151–165

    Google Scholar 

  • Li R, Murray AW (1991) Feedback control of mitosis in budding yeast. Cell 66: 519–531

    Google Scholar 

  • McAteer M, Donnan L, John PCL (1985) The timing of division inChlamydomonas. New Phytol 99: 41–56

    Google Scholar 

  • Meijer L (1995) Chemical inhibitors of cyclin-dependent kinases. Prog Cell Cycle Res 1: 351–363

    Google Scholar 

  • Mendenhall MD, Al-jumaily W, Nugroho YY (1995) The Cdc28 inhibitor p40SICI. Prog Cell Cycle Res 1: 173–185

    Google Scholar 

  • Millar JBA, McGowan CH, Lenaers G, Jones R, Russell P (1991) p80cdc25 mitotic inducer is the tyrosine phosphatase that activates p34cdc2 kinase in fission yeast EMBO J 10: 4301–4309

    Google Scholar 

  • Moreno S, Hayles J, Nurse P (1989) Regulation of p34cdc2 protein kinase during mitosis. Cell 58: 361–372

    Google Scholar 

  • Murray AW, Solomon ML, Kirschner MW (1989) The role of cyclin synthesis and degradation in the control of maturation promoting factor activity. Nature 339: 280–286

    Google Scholar 

  • Nigg EA (1995) Cyclin-dependent protein kinases; key regulators of the eukaryotic cell cycle. Bioessays 17: 471–480

    Google Scholar 

  • Niklas RB, Ward SC, Gorbsky GJ (1995) Kinetochore chemistry is sensitive to tension and may link mitotic forces to a cell cycle checkpoint. J Cell Biol 130: 929–939

    Google Scholar 

  • Nurse P (1990) Universal control mechanism regulating onset of Mphase. Nature 344: 503–508

    Google Scholar 

  • —, Bissett Y (1981) Gene required in g1 for commitment to cell cycle and in G2 for control of mitosis in fission yeast. Nature 292: 558–560

    Google Scholar 

  • Pringle JR (1981) The genetic approach to the study of the cell cycle. In: Zimmerman AM, Forer A (eds) Mitosis/cytokinesis. Academic Press, New York, pp 5–28

    Google Scholar 

  • Richardson HE, Wittenberg C, Cross F, Reed SI (1989) An essential G1 function for cyclin-like proteins in yeast. Cell 59: 1127–1133

    Google Scholar 

  • Roberts BT, Farr KA, Hoyt MA (1994) TheSaccharomyces cerevisiae checkpoint geneBUB1 encodes a novel protein kinase. Mol Cell Biol 14: 8282–8291

    Google Scholar 

  • Rollins MJ, Harper JDI, John PCL (1983) Synthesis of individual proteins, including tubulins and chloroplast membrane proteins, in synchronous cultures of the eukaryoteChlamydomonas reinhardtii: elimination of periodic changes in protein synthesis and enzyme activity under constant environmental conditions. J Gen Microbiol 129: 1899–1919

    Google Scholar 

  • Rudner AD, Murray AW (1996) The spindle assembly checkpoint. Curr Opin Cell Biol 8: 773–780

    Google Scholar 

  • Russell P, Nurse P (1987) The mitotic inducernim1 + functions in a regulatory network of protein kinase homologs controlling the inhibition of mitosis. Cell 49: 569–576

    Google Scholar 

  • Schibler MJ, Huang B (1991) Thecol R14 andcol R15 β-tubulin mutation inChlamydomonas reinhardtii confer altered sensitivities to microtubule inhibitors and herbicides by enhancing microtubule stability. J Cell Biol 113: 605–614

    Google Scholar 

  • Seufert W, Futcher B, Jentsch S (1995) Role of a ubiquitin-conjugating enzyme in degradation of S- and M-phase cyclins. Nature 373: 78–81

    Google Scholar 

  • Sherr CJ (1996) Cancer cell cycles. Science 274: 1672–1677

    Google Scholar 

  • Soni R, Carmichael JP, Shah ZH, Murray JAH (1995) A family of cyclin D homologs from plants differentially controlled by growth regulators and containing the conserved retinoblastoma protein interaction motif. Plant Cell 7: 85–103

    Google Scholar 

  • Staiger C, Doonan J (1993) Cell division in plants. Curr Opin Cell Biol 5: 226–231

    Google Scholar 

  • Stern B, Nurse P (1996) A quantitative model for the cdc2 control of S phase and mitosis in fission yeast. Trends Genet 12: 345–350.

    Google Scholar 

  • Sueoka N, Chiang K-S, Kates JR (1967) Deoxyribonucleic acid replication in meiosis ofChlamydomonas reinhardtii. I. Isotopic transfer experiments with a strain producing eight zoospores. J Mol Biol 25: 47–66

    Google Scholar 

  • Surana U, Amon A, Dowzer C, McGrew J, Byers B, Nasmyth K (1993) Destruction of the CDC28/CLB mitotic kinase is not required for the metaphase to anaphase transition in budding yeast. EMBO J 12: 1969–1978

    Google Scholar 

  • Suzuki DT, Griffiths AJF, Lewontin RC (1981) An introduction to genetic analysis, 2nd edn. WH Freeman, San Francisco

    Google Scholar 

  • Tomkinson AE, Tappe NJ, Friedberg EC (1992) DNA ligase 1 fromSaccharomyces cerevisiae: physical and biochemical characterisation of the CDC9 gene product. Biochemistry 31: 11762–11771

    Google Scholar 

  • Van't Hof J (1985) Control points within the cell cycle. In: Bryant JA, Francis D (eds) The cell division cycle in plants. Cambridge University Press, Cambridge, pp 1–13 (Society for Experimental Biology seminar series, vol 26)

    Google Scholar 

  • Weinert TA, Hartwell LH (1993) Cell cycle arrest of cdc mutants and specificity of theRAD9 checkpoint. Genetics 134: 63–80

    Google Scholar 

  • Wilson PJ, Forer A (1989) Acetylated α-tubulin in spermatogenic cells of the crane flyNephrotoma suturalis: kinetochore microtubules are selectively acetylated. Cell Motil Cytoskeleton 14: 237–250

    Google Scholar 

  • Zhang DH, Wadsworth P, Hepler PK (1990) Microtubule dynamics in living plant cells: confocal imaging of microinjected fluorescent brain tubulin. Proc Natl Acad Sci USA 87: 8820–8824

    Google Scholar 

  • Zhang K, Letham DSL, John PCL (1996) Cytokinin controls the cell cycle at mitosis by stimulating the tyrosine dephosphorylation and activation of p34cdc2-like H1 histone kinase. Planta 200: 2–12

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wu, L., Hepler, P.K. & John, P.C.L. Themet1 mutation inChlamydomonas reinhardtii causes arrest at mitotic metaphase with persisting p34cdc2-like H1 histone kinase activity that can promote mitosis when injected into higher-plant cells. Protoplasma 199, 135–150 (1997). https://doi.org/10.1007/BF01294502

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01294502

Keywords

Navigation