Phosphatidylinositol-4-phosphate 5-kinase regulates fission yeast cell integrity through a phospholipase C-mediated protein kinase C-independent pathway

J Biol Chem. 2005 Jul 29;280(30):27561-8. doi: 10.1074/jbc.M502660200. Epub 2005 May 27.

Abstract

Fission yeast its3-1 mutant is an allele of the essential gene its3+ that encodes a phosphatidylinositol-4-phosphate 5-kinase (PIP5K) that produces phosphatidylinositol 4,5-bisphosphate. We found that the its3-1 mutant is sensitive to micafungin, a (1,3)-beta-D-glucan synthase inhibitor, suggesting a cell wall integrity defect. Consistently, its3-1 mutation caused synthetic lethality with a (1,3)-beta-D-glucan synthase mutant, bgs1-i2, and its3-1 mutant cells showed aberrant localization of green fluorescent protein-Bgs1. Similar aberrant localization of green fluorescent protein-tagged Rgf1, a putative phosphatidylinositol 4,5-bisphosphate-binding guanine nucleotide exchange factor for Rho protein, in its3-1 mutants was observed, suggesting a defective Rgf1/Rho pathway. To unravel the molecular mechanism(s), putative downstream components of PIP5K signaling were analyzed. Unexpectedly, overexpression of phospholipase C (Plc1), but not that of protein kinase C (PKC; Pck1 and Pck2), suppressed the phenotypes of the its3-1 mutant. These findings indicate that PKCs are not involved in the suppression, and further analysis revealed that PKCs are not downstream of Plc1 in fission yeast. Also, the enzymatic activity of Plc1 is essential for the suppression of the phenotypes and for the viability of the its3-1 mutant. These findings suggest that Its3 PIP5K regulates cell integrity through a Plc1-mediated PKC-independent pathway, in addition to the Rho/PKC pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Cell Wall / drug effects
  • Cell Wall / metabolism
  • DNA Primers / metabolism
  • Echinocandins
  • Gene Deletion
  • Gene Expression Regulation, Fungal*
  • Glucosyltransferases / genetics
  • Green Fluorescent Proteins / metabolism
  • Lipopeptides
  • Lipoproteins / pharmacology
  • Micafungin
  • Models, Biological
  • Mutagenesis, Site-Directed
  • Mutation*
  • Peptides, Cyclic / pharmacology
  • Phenotype
  • Phosphatidylinositol 4,5-Diphosphate / metabolism
  • Phosphotransferases (Alcohol Group Acceptor) / physiology*
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Kinase C / metabolism*
  • Schizosaccharomyces
  • Signal Transduction
  • Temperature
  • Type C Phospholipases / metabolism*

Substances

  • DNA Primers
  • Echinocandins
  • Lipopeptides
  • Lipoproteins
  • Peptides, Cyclic
  • Phosphatidylinositol 4,5-Diphosphate
  • Green Fluorescent Proteins
  • Glucosyltransferases
  • 1,3-beta-glucan synthase
  • Phosphotransferases (Alcohol Group Acceptor)
  • 1-phosphatidylinositol-4-phosphate 5-kinase
  • Protein Kinase C
  • Type C Phospholipases
  • Micafungin