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PMID: 8196614 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Two types of RAS mutants that dominantly interfere with activators of RAS.

Molecular and cellular biology ·Vol. 14 ·No. 6 ·1994-06-00 ·Pages 3707-18

Jung V, Wei W, Ballester R, Camonis J, Mi S, Van Aelst L, Wigler M, Broek D

Abstract

In the fission yeast Schizosaccharomyces pombe, ras1 regulates both sexual development (conjugation and sporulation) and cellular morphology. Two types of dominant interfering mutants were isolated in a genetic screen for ras1 mutants that blocked sexual development. The first type of mutation, at Ser-22, analogous to the H-rasAsn-17 mutant (L. A. Feig and G. M. Cooper, Mol. Cell. Biol. 8:3235-3243, 1988), blocked only conjugation, whereas a second type of mutation, at Asp-62, interfered with conjugation, sporulation, and cellular morphology. Analogous mutations at position 64 of Saccharomyces cerevisiae RAS2 or position 57 of human H-ras also resulted in dominant interfering mutants that interfered specifically and more profoundly than mutants of the first type with RAS-associated pathways in both S. pombe or S. cerevisiae. Genetic evidence indicating that both types of interfering mutants function upstream of RAS is provided. Biochemical evidence showing that the mutants are altered in their interaction with the CDC25 class of exchange factors is presented. We show that both H-rasAsn-17 and H-rasTyr-57, compared with wild-type H-ras, are defective in their guanine nucleotide-dependent release from human cdc25 and that this defect is more severe for the H-rasTyr-57 mutant. Such a defect would allow the interfering mutants to remain bound to, thereby sequestering RAS exchange factors. The more severe interference phenotype of this novel interfering mutant suggests that it functions by titrating out other positive regulators of RAS besides those encoded by ste6 and CDC25.

Related Genes
H-ras RAS RAS2 ras1
MeSH Terms
Base Sequence Cloning, Molecular Conjugation, Genetic Fungal Proteins/biosynthesis,genetics Genes, Fungal Genes, ras Genotype Molecular Sequence Data Mutagenesis, Site-Directed Oligodeoxyribonucleotides Plasmids Restriction Mapping Saccharomyces cerevisiae/genetics,growth & development,physiology Saccharomyces cerevisiae Proteins Schizosaccharomyces/genetics,growth & development,physiology Schizosaccharomyces pombe Proteins Spores, Fungal/genetics,physiology ras Proteins
Chemicals
Fungal Proteins Oligodeoxyribonucleotides Saccharomyces cerevisiae Proteins Schizosaccharomyces pombe Proteins RAS1 protein, S cerevisiae RAS2 protein, S cerevisiae Ras1 protein, S pombe ras Proteins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Jung V
Cold Spring Harbor Laboratory, New York 11724.
Wei W
Ballester R
Camonis J
Mi S
Van Aelst L
Wigler M
Broek D
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.6.3707-3718.1994
Published
1994-06-00
Pages
3707-18
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358738
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.14.6.3707-3718.1994 PMC Full Text View journal details
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