Molecular and Functional Characterization of a Mutant Allele of the Mitogen-activated Protein-kinase Gene SLT2(MPK1) Rescued from Yeast Autolytic Mutants

Overview

Journal Curr Genet

Specialty Genetics

Date 1996 May 1

PMID 8662190

Citations 20

Authors

H Martin

M C Castellanos

R Cenamor

M Sanchez

M Molina

C Nombela

Affiliations

Soon will be listed here.

Abstract

We have further characterized the functionality of the Saccharomyces cerevisiae gene SLT2(MPK1), coding for a MAP-kinase homolog essential for cell integrity, which is involved in the Pkc1p signalling pathway. This gene was isolated on the basis of its capacity to complement the thermosensitive-autolytic, osmotic-remediable phenotype of lyt2 mutants. Both slt2delta and lyt2 mutants displayed a caffeine-sensitive phenotype consisting of cell lysis that was not dependent on temperature. Caffeine concentrations affecting the growth of these mutant strains were dependent on the genetic background, the SSD1 allele being very significant in this regard. The SLT2 allele of several lyt2 strains was both rescued and amplified by PCR. The recovered allele was shown to be non-functional as it could not complement the lytic phenotype of both deletion (slt2delta) and lyt2 strains. After nucleotide sequencing of the recovered allele, we found that the defect of lyt2 mutants consists in a substitution of an aspartic acid for a glycine at position 35 of the amino-acid sequence of Slt2p. Gly35 is the third glycine of a glycine cluster (Gly-X-Gly-X-X-Gly), a conserved region in protein kinases and other nucleotide-binding proteins. Keywords Yeast middle dot SLT2 middle dot MAP-kinase middle dot Caffeine

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