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Disruption of six novel yeast genes located on chromosome II reveals one gene essential for vegetative growth and two required for sporulation and conferring hypersensitivity to various chemicals
Author(s) -
Kucharczyk R.,
Gromadka R.,
Migdalski A.,
Słonimski P. P.,
Rytka J.
Publication year - 1999
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(199907)15:10b<987::aid-yea403>3.0.co;2-5
Subject(s) - biology , gene , mutant , genetics , phenotype , orfs , chromosome , ploidy , genetic screen , open reading frame , microbiology and biotechnology , peptide sequence
A PCR‐based method for targeted gene deletion by kanMX4 module was used to construct complete deletion mutants of six individual open reading frames from chromosome II: YBR128c , YBR131w , YBR133c , YBR137w , YBR138c and YBR142w . The ORFs were deleted in two diploid strains, FY1679 and W303. Sporulation and tetrad analysis revealed that only one ORF, YBR142w , encoding a putative DEAD‐box RNA helicase, is an essential gene. A systematic phenotypic analysis of the deleted mutants was carried out. Homozygous diploids ybr128cΔ /ybr128cΔ and ybr131wΔ /ybr131wΔ did not sporulate. The ybr131cΔ mutant whether haploid or homozygous diploid, in addition displayed an increased sensitivity to C affeine, C alcium and Z inc, and to emphasize this phenotype we named the gene CCZ1. ORF YBR133c was independently reported by others as H istone S ynthetic L ethal ( HSL7 ) (Ma et al ., 1996). We found that the aberrant morphology characteristic for ybr133cΔ ( hsl7Δ ) cells was observed in W303 but not in FY1679 genetic background. Furthermore, we observed that deletion of YBR133c had a pleiotropic effect under a wide range of conditions, including increased sensitivity to calcium, caffeine, calcofluor white, vanadate and verapamil. The effects of the deletion were reinforced in W303 background. We found no phenotypic effects of the two remaining deletions, ybr137wΔ and ybr138cΔ . Copyright © 1999 John Wiley & Sons, Ltd.