Identification of a mid-anaphase checkpoint in budding yeast

J Cell Biol. 1997 Jan 27;136(2):345-54. doi: 10.1083/jcb.136.2.345.

Abstract

Activation of a facultative, dicentric chromosome provides a unique opportunity to introduce a double strand DNA break into a chromosome at mitosis. Time lapse video enhanced-differential interference contrast analysis of the cellular response upon dicentric activation reveals that the majority of cells initiates anaphase B, characterized by pole-pole separation, and pauses in mid-anaphase for 30-120 min with spindles spanning the neck of the bud before completing spindle elongation and cytokinesis. The length of the spindle at the delay point (3-4 microm) is not dependent on the physical distance between the two centromeres, indicating that the arrest represents surveillance of a dicentric induced aberration. No mid-anaphase delay is observed in the absence of the RAD9 checkpoint gene, which prevents cell cycle progression in the presence of damaged DNA. These observations reveal RAD9-dependent events well past the G2/M boundary and have considerable implications in understanding how chromosome integrity and the position and state of the mitotic spindle are monitored before cytokinesis.

Publication types

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

MeSH terms

  • Anaphase*
  • Cell Cycle Proteins*
  • Cell Nucleus / physiology
  • Cell Nucleus / ultrastructure
  • Chromatids / physiology
  • Chromosomes, Fungal / physiology*
  • Chromosomes, Fungal / ultrastructure
  • Fluorescent Antibody Technique
  • Fungal Proteins / genetics*
  • Genes, Fungal
  • In Situ Hybridization, Fluorescence
  • Kinetics
  • Microscopy, Interference
  • Microscopy, Video
  • Mitosis
  • Morphogenesis
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / genetics
  • Spindle Apparatus / physiology*
  • Spindle Apparatus / ultrastructure

Substances

  • Cell Cycle Proteins
  • Fungal Proteins
  • rad9 protein