Current Biology
Volume 3, Issue 1, January 1993, Pages 13-26
Journal home page for Current Biology

Paper
Mitotic regulation of protein phosphatases by the fission yeast sds22 protein

https://doi.org/10.1016/0960-9822(93)90140-JGet rights and content

Abstract

Background: Cell cycle progression requires the activity of protein kinases and phosphatases at critical points in the cell cycle in all eukaryotes. We have previously reported that the dis2+ and sds2+ genes of fission yeast encode redundant catalytic subunits of a type 1-like protein phosphatase. The sds22+ gene was shown to be essential for cell viability and to interact genetically with dis2+ and sds21+.

Results: Here we show by immunoprecipitation that the sds22 protein physically interacts with the dis2 and sds21 proteins, and that sds22-associated phosphatase activity has altered substrate specificity, The loss of sds22 function by a temperature sensitive mutation leads to cell cycle arrest at mid-mitosis, at which point cdc2-dependent histone Hl kinase activity is high while sds22-dependent H1 phosphatase activity is low. To examine the unusual properties of sds22 protein structure, we analyzed a collection of sds22 deletion and point mutants by a variety of functional criteria.

Conclusion: We propose that sds22 is a regulatory subunit of the dis2/sds21 phosphatase catalytic subunits and that sds22-bound phosphatase carries a key phosphatase activity essential for the progression from metaphase to anaphase. Mutational analysis indicates that dis2/sds21 interacts with the central repetitive domain of sds22, while the C-terminal and central regions of sds22 may be involved in subcellular targeting and the N-terminus is important for stability.

References (44)

  • S. Moreno et al.

    Molecular genetic analysis of fission yeast Scbizosambaromyces pombe

    Meth Enz

    (1991)
  • M.J. Solomon et al.

    Cyclin activation of p34cdc2/

    Cell

    (1990)
  • P. NursE

    Universal control mechanism regulating onset of M-phase

    Nature

    (1990)
  • J.B.A. Miliar et al.

    The cdc25 M-phase inducer: an unconventional protein phosphatase

    Cell

    (1992)
  • H. Ohkura et al.

    The fission yeast dis2 gene required for chromosome disjoining encodes one of two putative type-1 protein phosphatases

    Cell

    (1989)
  • M-A. Felix et al.

    Cdc2 H1 kinase is negatively regulated by a type 2A phosphatase in the Xenopus early embryonic cell cycle: evidence from the effects of okadaic acid

    EMBO J

    (1990)
  • K. Yamashita et al.

    Okadaic acid, a potent inhibitor of type 1 and type 2A phosphatases, activates cdc2/Hl kinase and transiently induces a premature mitosis-like state in BHK21 cells

    EMBO J

    (1990)
  • N. Kinoshita et al.

    Sister-chromatid separation and protein dephosphorylation in mitosis

  • P. Cohen

    The structure and regulation of protein phosphatases

    Annu Rev Biocbem

    (1989)
  • P. Stralfors et al.

    The protein phosphatases involved in cellular regulation: Purification and characterisation of the glycogen-bound form of protein phosphatase-1 from rabbit skeletal muscle

    Eur J Biochem

    (1985)
  • O.A.K. Chisholm et al.

    The myosin-bound form of protein phosphatase 1 (PP-1M) is the enzyme that dephosphorylates native myosin in skeletal and cardiac muscles

    Biochem Biophys Acta

    (1988)
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    Present address: Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, CO 80309, USA.

    Present address: Department of Biochemistry and Biophysics, University of California School of Medicine, San Francisco, CA 94143, USA.

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