Spatial control of Cdc42 activation determines cell width in fission yeast

Mol Biol Cell. 2011 Oct;22(20):3801-11. doi: 10.1091/mbc.E11-01-0057. Epub 2011 Aug 17.

Abstract

The fission yeast Schizosaccharomyces pombe is a rod-shaped cell that grows by linear extension at the cell tips, with a nearly constant width throughout the cell cycle. This simple geometry makes it an ideal system for studying the control of cellular dimensions. In this study, we carried out a near-genome-wide screen for mutants wider than wild-type cells. We found 11 deletion mutants that were wider; seven of the deleted genes are implicated in the control of the small GTPase Cdc42, including the Cdc42 guanine nucleotide exchange factor (GEF) Scd1 and the Cdc42 GTPase-activating protein (GAP) Rga4. Deletions of rga4 and scd1 had additive effects on cell width, and the proteins localized independently of one another, with Rga4 located at the cell sides and Scd1 at the cell tips. Activated Cdc42 localization is altered in rga4Δ, scd1Δ, and scd2Δ mutants. Delocalization and ectopic retargeting experiments showed that the localizations of Rga4 and Scd1 are crucial for their roles in determining cell width. We propose that the GAP Rga4 and the GEF Scd1 establish a gradient of activated Cdc42 within the cellular tip plasma membrane, and it is this gradient that determines cell growth-zone size and normal cell width.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Division / genetics
  • Cell Enlargement
  • Cell Shape
  • Cell Size
  • Cytoskeleton / genetics
  • Cytoskeleton / metabolism*
  • GTPase-Activating Proteins / deficiency
  • GTPase-Activating Proteins / genetics*
  • Gene Deletion
  • Gene Expression Regulation, Fungal
  • Guanine Nucleotide Exchange Factors / deficiency
  • Guanine Nucleotide Exchange Factors / genetics*
  • Microscopy, Fluorescence
  • Microscopy, Interference
  • Organisms, Genetically Modified / genetics
  • Organisms, Genetically Modified / growth & development
  • Organisms, Genetically Modified / metabolism*
  • Plasmids
  • Protein Binding
  • Schizosaccharomyces / cytology
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / growth & development
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins / genetics*
  • Signal Transduction / genetics*
  • Transduction, Genetic
  • cdc42 GTP-Binding Protein, Saccharomyces cerevisiae / deficiency
  • cdc42 GTP-Binding Protein, Saccharomyces cerevisiae / genetics*

Substances

  • GTPase-Activating Proteins
  • Guanine Nucleotide Exchange Factors
  • Rga4 protein, S pombe
  • Schizosaccharomyces pombe Proteins
  • scd1 protein, S pombe
  • cdc42 GTP-Binding Protein, Saccharomyces cerevisiae