Dynamic positioning of mitotic spindles in yeast: role of microtubule motors and cortical determinants

Mol Biol Cell. 2000 Nov;11(11):3949-61. doi: 10.1091/mbc.11.11.3949.

Abstract

In the budding yeast Saccharomyces cerevisiae, movement of the mitotic spindle to a predetermined cleavage plane at the bud neck is essential for partitioning chromosomes into the mother and daughter cells. Astral microtubule dynamics are critical to the mechanism that ensures nuclear migration to the bud neck. The nucleus moves in the opposite direction of astral microtubule growth in the mother cell, apparently being "pushed" by microtubule contacts at the cortex. In contrast, microtubules growing toward the neck and within the bud promote nuclear movement in the same direction of microtubule growth, thus "pulling" the nucleus toward the bud neck. Failure of "pulling" is evident in cells lacking Bud6p, Bni1p, Kar9p, or the kinesin homolog, Kip3p. As a consequence, there is a loss of asymmetry in spindle pole body segregation into the bud. The cytoplasmic motor protein, dynein, is not required for nuclear movement to the neck; rather, it has been postulated to contribute to spindle elongation through the neck. In the absence of KAR9, dynein-dependent spindle oscillations are evident before anaphase onset, as are postanaphase dynein-dependent pulling forces that exceed the velocity of wild-type spindle elongation threefold. In addition, dynein-mediated forces on astral microtubules are sufficient to segregate a 2N chromosome set through the neck in the absence of spindle elongation, but cytoplasmic kinesins are not. These observations support a model in which spindle polarity determinants (BUD6, BNI1, KAR9) and cytoplasmic kinesin (KIP3) provide directional cues for spindle orientation to the bud while restraining the spindle to the neck. Cytoplasmic dynein is attenuated by these spindle polarity determinants and kinesin until anaphase onset, when dynein directs spindle elongation to distal points in the mother and bud.

Publication types

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

MeSH terms

  • Anaphase / physiology
  • Base Sequence
  • Cell Surface Extensions / genetics
  • Cytoplasm / genetics
  • Cytoplasm / metabolism
  • Cytoplasm / ultrastructure
  • Dyneins / metabolism
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Kinesins
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism
  • Microtubules / metabolism*
  • Microtubules / ultrastructure
  • Molecular Sequence Data
  • Mutation
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Saccharomyces cerevisiae Proteins*
  • Spindle Apparatus / metabolism
  • Spindle Apparatus / ultrastructure*
  • Yeasts / genetics
  • Yeasts / metabolism
  • Yeasts / ultrastructure*

Substances

  • BUD6 protein, S cerevisiae
  • Bni1 protein, S cerevisiae
  • Fungal Proteins
  • KAR9 protein, S cerevisiae
  • KIP3 protein, S cerevisiae
  • Microfilament Proteins
  • Microtubule-Associated Proteins
  • Nuclear Proteins
  • Saccharomyces cerevisiae Proteins
  • Dyneins
  • Kinesins