Total in vitro maturation of the Saccharomyces cerevisiae a-factor lipopeptide mating pheromone

Academic Article

Abstract

  • The a-factor mating pheromone, produced by Saccharomyces cerevisiae a haploid cells, is posttranslationally modified in a manner analogous to that of the rasproto-oncogene product. A consensus C-terminal amino acid sequence, -CAAX (C is cysteine, A is aliphatic amino acid, and X is any amino acid), is the target of these modifications, which include isoprenylation (essential for Ras function), proteolysis of the -AAX sequence, and carboxy methyl esterification. Recently, the RAM DPR1 gene product was shown to be a component of the activity responsible for isoprenylation of both Ras and a-factor. In this report, we present an in vitro assay which not only detects a-factor isoprenylation, but also proteolysis and carboxy methyl esterification, and directly demonstrates, biochemically, the order of these processing events. This a-factor maturation assay may prove useful for screening agents which block any of the steps involved in the post-translational modification of the a-factor and Ras -CAAX sequences. Such agents would be potential anti-Ras-related cancer therapeutic drugs. © 1990.
  • Authors

    Digital Object Identifier (doi)

    Author List

  • Marcus S; Caldwell GA; Xue CB; Naider F; Becker JM
  • Start Page

  • 1310
  • End Page

  • 1316
  • Volume

  • 172
  • Issue

  • 3