Molecular cloning of CWP1: a gene encoding a Saccharomyces cerevisiae cell wall protein solubilized with Rarobacter faecitabidus protease I

J Biochem. 1995 Aug;118(2):302-11. doi: 10.1093/oxfordjournals.jbchem.a124907.

Abstract

A yeast cell wall glycoprotein with a molecular weight of 40,000, named gp40, was solubilized from SDS-extracted cell wall of Saccharomyces cerevisiae by incubation with Rarobacter faecitabidus protease I, which is a yeast-lytic enzyme. Based on its amino acid sequence, we cloned and sequenced the gene encoding the precursor of gp40, named CWP1; cell wall protein gene. The DNA sequence of the CWP1 gene was identical to YKL443, an open reading frame identified in a genome sequencing program for yeast chromosome XI. This gene encoded a serine-rich protein of 239 amino acids with a molecular weight of 24,267. The presence of hydrophobic sequences in the N- and C-termini of the CWP1 protein suggests that it is secreted as a glycosylphosphatidylinositol-anchored protein and is subsequently integrated into the cell wall. Since a gene disruption experiment showed no growth defect, the CWP1 gene is not essential for growth. Mutant CWP1 protein deficient in the C-terminal hydrophobic sequence was secreted into the culture medium, not anchored to the cell wall, thereby indicating that this hydrophobic sequence plays a crucial role in anchoring to the cell wall. Homology between the CWP1 protein and TIP1 family of cold shock proteins suggests that they belong to a new family of cell wall proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacteria / enzymology*
  • Base Sequence
  • Cell Wall / metabolism*
  • Chromosome Mapping
  • Cloning, Molecular
  • DNA, Complementary
  • Endopeptidases / metabolism*
  • Gram-Positive Asporogenous Rods / enzymology
  • Membrane Glycoproteins / genetics*
  • Molecular Sequence Data
  • Peptide Fragments / genetics*
  • Protein Precursors / genetics
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins*
  • Transcription, Genetic

Substances

  • CWP1 protein, S cerevisiae
  • DNA, Complementary
  • Membrane Glycoproteins
  • Peptide Fragments
  • Protein Precursors
  • Saccharomyces cerevisiae Proteins
  • Endopeptidases

Associated data

  • GENBANK/D37975