Characterization of canavanine-resistance of cat1 and vhc1 deletions and a dominant any1 mutation in fission yeast

PLoS One. 2022 May 31;17(5):e0269276. doi: 10.1371/journal.pone.0269276. eCollection 2022.

Abstract

Positive and counter-selectable markers have been successfully integrated as a part of numerous genetic assays in many model organisms. In this study, we investigate the mechanism of resistance to arginine analog canavanine and its applicability for genetic selection in Schizosaccharomyces pombe. Deletion of both the arginine permease gene cat1 and SPBC18H10.16/vhc1 (formerly mistakenly called can1) provides strong drug resistance, while the single SPBC18H10.16/vhc1 deletion does not have an impact on canavanine resistance. Surprisingly, the widely used can1-1 allele does not encode for a defective arginine permease but rather corresponds to the any1-523C>T allele. The strong canavanine-resistance conferred by this allele arises from an inability to deposit basic amino acid transporters on the cellular membrane. any1-523C>T leads to reduced post-translational modifications of Any1 regulated by the Tor2 kinase. We also demonstrate that any1-523C>T is a dominate allele. Our results uncover the mechanisms of canavanine-resistance in fission yeast and open the opportunity of using cat1, vhc1 and any1 mutant alleles in genetic assays.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Alleles
  • Amino Acid Transport Systems* / genetics
  • Amino Acid Transport Systems* / metabolism
  • Arginine / metabolism
  • Arrestins* / genetics
  • Arrestins* / metabolism
  • Canavanine / metabolism
  • Mutation
  • Schizosaccharomyces pombe Proteins* / genetics
  • Schizosaccharomyces pombe Proteins* / metabolism
  • Schizosaccharomyces* / genetics
  • Schizosaccharomyces* / metabolism
  • Symporters* / genetics
  • Symporters* / metabolism

Substances

  • Amino Acid Transport Systems
  • Any1 protein, S pombe
  • Arrestins
  • Cat1 protein, S pombe
  • Schizosaccharomyces pombe Proteins
  • Symporters
  • Canavanine
  • Arginine