Drosophila grapes/CHK1 mutants are defective in cyclin proteolysis and coordination of mitotic events

Curr Biol. 1999 Aug 26;9(16):919-22. doi: 10.1016/s0960-9822(99)80399-6.

Abstract

The Drosophila grapes (grp) gene, which encodes a homolog of the Schizosaccharomyces pombe Chk1 kinase, provides a cell-cycle checkpoint that delays mitosis in response to inhibition of DNA replication [1]. Grp is also required in the undisturbed early embryonic cycles: in its absence, mitotic abnormalities appear in cycle 12 and chromosomes fail to fully separate in subsequent cycles [2] [3]. In other systems, Chk1 kinase phosphorylates and suppresses the activity of Cdc25 phosphatase: the resulting failure to remove inhibitory phosphate from cyclin-dependent kinase 1 (Cdk1) prevents entry into mitosis [4] [5]. Because in Drosophila embryos Cdk1 lacks inhibitory phosphate during cycles 11-13 [6], it is not clear that known actions of Grp/Chk1 suffice in these cycles. We found that the loss of grp compromised cyclin A proteolysis and delayed mitotic disjunction of sister chromosomes. These defects occurred before previously reported grp phenotypes. We conclude that Grp activates cyclin A degradation, and functions to time the disjunction of chromosomes in the early embryo. As cyclin A destruction is required for sister chromosome separation [7], a failure in Grp-promoted cyclin destruction can also explain the mitotic phenotype. The mitotic failure described previously for cycle 12 grp embryos might be a more severe form of the phenotypes that we describe in earlier embryos and we suggest that the underlying defect is reduced degradation of cyclin A.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Anaphase / genetics
  • Animals
  • Blotting, Western
  • Cell Cycle / drug effects
  • Cell Cycle / genetics
  • Checkpoint Kinase 1
  • Cyclin A / metabolism*
  • Cycloheximide / pharmacology
  • DNA / analysis
  • Drosophila / cytology
  • Drosophila / enzymology
  • Drosophila / genetics*
  • Drosophila / growth & development*
  • Drosophila Proteins
  • Microscopy, Fluorescence
  • Protein Kinases / genetics
  • Protein Kinases / physiology*
  • Protein Synthesis Inhibitors / pharmacology
  • Schizosaccharomyces pombe Proteins

Substances

  • Cyclin A
  • Drosophila Proteins
  • Protein Synthesis Inhibitors
  • Schizosaccharomyces pombe Proteins
  • DNA
  • Cycloheximide
  • Protein Kinases
  • Checkpoint Kinase 1
  • Chk1 protein, S pombe
  • grp protein, Drosophila