Pck2 association with the plasma membrane and efficient response of the cell integrity pathway require regulation of PI4P homeostasis by exomer

Open Biol. 2024 Nov;14(11):240101. doi: 10.1098/rsob.240101. Epub 2024 Nov 13.

Abstract

Exomer is a protein complex that facilitates trafficking between the Golgi and the plasma membrane (PM). Schizosaccharomyces pombe exomer is composed of Cfr1 and Bch1, and we have found that full activation of the cell integrity pathway (CIP) in response to osmotic stress requires exomer. In the wild-type, the CIP activators Rgf1 (Rho1 GEF) and Pck2 (PKC homologue) and the MEK kinase Mkh1 localize in the PM, internalize after osmotic shock and re-localize after adaptation. This re-localization is inefficient in exomer mutants. Overexpression of the PM-associated 1-phosphatidylinositol 4-kinase stt4+, and deletion of the nem1+ phosphatase suppress the defects in Pck2 dynamics in exomer mutants, but not their defect in CIP activation, demonstrating that exomer regulates CIP in additional ways. Exomer mutants accumulate PI4P in the TGN, and increasing the expression of the Golgi-associated 1-phosphatidylinositol 4-kinase pik1+ suppresses their defect in Pck2 dynamics. These findings suggest that efficient PI4P transport from the Golgi to the PM requires exomer. Mutants lacking clathrin adaptors are defective in CIP activation, but not in Pck2 dynamics or in PI4P accumulation in the Golgi. Hence, traffic from the Golgi regulates CIP activation, and exomer participates in this regulation through an exclusive mechanism.

Keywords: cell integrity pathway; fission yeast; phosphatidylinositol 4-phosphate; phospholipids; stress response; trans-Golgi network.

MeSH terms

  • Cell Membrane* / metabolism
  • Golgi Apparatus / metabolism
  • Homeostasis*
  • Mutation
  • Osmotic Pressure
  • Phosphatidylinositol Phosphates / metabolism
  • Protein Transport
  • Schizosaccharomyces pombe Proteins* / genetics
  • Schizosaccharomyces pombe Proteins* / metabolism
  • Schizosaccharomyces* / genetics
  • Schizosaccharomyces* / metabolism
  • Signal Transduction

Substances

  • Schizosaccharomyces pombe Proteins
  • phosphatidylinositol 4-phosphate
  • Phosphatidylinositol Phosphates