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- Title
Isolation of Synthetic Lethal Mutants of rasl in S chizosaccharomyces pombe
- Authors
Kyung-Sook Chung; Kyu-Won Kim; Hyang-Sook Yoo
- Abstract
Ras proteins are membrane-associated guanine nucleotide-binding proteins that serve as molecular switches for signal transduction pathways in a diverse array of organisms. Various cellular factors are known to interact with Ras proteins. In order to find the novel cellular factors that are associated with Ras function, we have constructed synthetic lethal mutants of the ras1+ gene in Schizosaccharomyces pombe and used them to identify the genes that are functionally dependent on the Ras1. We first constructed S. pombe strains in which chromosomal ras1+ gene is placed under the nmt1 promoter that is regulated by thiamine. This strain shows rasr phenotype in the absence of thiamine, whereas it shows ras1 - phenotype in the presence of thiamine. Second, we mutated the constructed strains with ultraviolet light (UV) and selected two synthetic lethal mutants that could not grow when Rasl function was repressed (ras1). One of the mutants, KSC3, showed a swollen cell shape, aberrant deposition of septum materials, and aberrant nuclei. The other mutant, KSC4, showed sensitivity to hyper-osmolarity when Rasl function is absent. These mutants, however, grow normally when Ras1 is expressed (ras1+). These two novel synthetic lethal mutants of rasl provide the means to isolate the corresponding genes that function in association with Rasl in S. pombe. Screening of a genomic library of S. pombe complementing the mutant phenotype allowed us to identify several novel genes associated with Rasl of S. pombe.
- Publication
Molecules & Cells (Springer Nature), 1997, Vol 7, Issue 6, p800
- ISSN
1016-8478
- Publication type
Article
- DOI
10.1016/s1016-8478(23)13382-6