A systematic screen for morphological abnormalities during fission yeast sexual reproduction identifies a mechanism of actin aster formation for cell fusion

PLoS Genet. 2017 Apr 14;13(4):e1006721. doi: 10.1371/journal.pgen.1006721. eCollection 2017 Apr.

Abstract

In non-motile fungi, sexual reproduction relies on strong morphogenetic changes in response to pheromone signaling. We report here on a systematic screen for morphological abnormalities of the mating process in fission yeast Schizosaccharomyces pombe. We derived a homothallic (self-fertile) collection of viable deletions, which, upon visual screening, revealed a plethora of phenotypes affecting all stages of the mating process, including cell polarization, cell fusion and sporulation. Cell fusion relies on the formation of the fusion focus, an aster-like F-actin structure that is marked by strong local accumulation of the myosin V Myo52, which concentrates secretion at the fusion site. A secondary screen for fusion-defective mutants identified the myosin V Myo51-associated coiled-coil proteins Rng8 and Rng9 as critical for the coalescence of the fusion focus. Indeed, rng8Δ and rng9Δ mutant cells exhibit multiple stable dots at the cell-cell contact site, instead of the single focus observed in wildtype. Rng8 and Rng9 accumulate on the fusion focus, dependent on Myo51 and tropomyosin Cdc8. A tropomyosin mutant allele, which compromises Rng8/9 localization but not actin binding, similarly leads to multiple stable dots instead of a single focus. By contrast, myo51 deletion does not strongly affect fusion focus coalescence. We propose that focusing of the actin filaments in the fusion aster primarily relies on Rng8/9-dependent cross-linking of tropomyosin-actin filaments.

MeSH terms

  • Actin Cytoskeleton / genetics
  • Actin Cytoskeleton / metabolism
  • Actins / genetics
  • Amino Acid Sequence / genetics
  • Cell Cycle Proteins / genetics*
  • Cytoskeleton / genetics
  • Cytoskeleton / metabolism
  • Myosin Type V / genetics*
  • Myosin Type V / metabolism
  • Myosins / genetics*
  • Phenotype
  • Protein Binding
  • Reproduction / genetics
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces pombe Proteins / genetics*
  • Sequence Deletion

Substances

  • Actins
  • Cdc8 protein, S pombe
  • Cell Cycle Proteins
  • Rng8 protein, S pombe
  • Rng9 protein, S pombe
  • Schizosaccharomyces pombe Proteins
  • Myo52 protein, S pombe
  • Myosin Type V
  • Myo51 protein, S pombe
  • Myosins

Grants and funding

This work was supported by an ERC Consolidator grant (CellFusion) and a Swiss National Science Foundation (31003A_155944) to SGM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.