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Growth-based Determination and Biochemical Confirmation of Genetic Requirements for Protein Degradation in Saccharomyces cerevisiae
10:57

Growth-based Determination and Biochemical Confirmation of Genetic Requirements for Protein Degradation in Saccharomyces cerevisiae

Published on: February 16, 2015

Two distinct genes encode small isoproteolipids affecting plasma membrane H(+)-ATPase activity of Saccharomyces

C Navarre1, P Catty, S Leterme

  • 1Unité de Biochimie Physiologique, Université Catholique de Louvain, Louvain-la-Neuve, Belgium.

The Journal of Biological Chemistry
|August 19, 1994
PubMed

Insights

Researchers identified PMP2, a second proteolipid gene in yeast plasma membranes. Deleting both PMP genes reduced H(+)-ATPase activity, suggesting these proteolipids regulate enzyme function.

Area of Science:

  • Molecular and Cellular Biology
  • Biochemistry
  • Yeast Genetics

Background:

  • The plasma membrane H(+)-ATPase is crucial for yeast cell function.
  • Proteolipids are membrane-associated proteins with roles in cellular processes.
  • PMP1 was previously identified as a proteolipid associated with the yeast plasma membrane H(+)-ATPase.

Purpose of the Study:

  • To identify and characterize additional proteolipid genes in Saccharomyces cerevisiae.
  • To investigate the functional relationship between proteolipids and the plasma membrane H(+)-ATPase.

Main Methods:

  • Gene identification using hybridization with a PMP1 probe.
  • DNA sequencing to determine gene identity and homology.
  • Chloroform/methanol extraction to isolate proteolipids.
  • Enzyme kinetic analysis (Vmax determination) of H(+)-ATPase activity.

Main Results:

  • A second Saccharomyces cerevisiae plasma membrane proteolipid gene, PMP2, was identified.
  • PMP2 shares 92% sequence identity with PMP1, encoding a 43-amino acid polypeptide.
  • The PMP1 and PMP2 genes are co-expressed, and deletion of one does not affect the other's transcription.
  • Yeast strains lacking both PMP genes exhibited reduced Vmax for plasma membrane H(+)-ATPase activity.

Conclusions:

  • Two distinct but related proteolipid genes, PMP1 and PMP2, exist in yeast.
  • These proteolipids play a regulatory role in the activity of the yeast plasma membrane H(+)-ATPase.
  • The absence of both PMP genes significantly impairs the enzyme's maximal catalytic rate.

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