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酿酒酵母正常生长所需的新基因SUA5的分离与鉴定。

Isolation and characterization of SUA5, a novel gene required for normal growth in Saccharomyces cerevisiae.

作者信息

Na J G, Pinto I, Hampsey M

机构信息

Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, Shreveport 71130.

出版信息

Genetics. 1992 Aug;131(4):791-801. doi: 10.1093/genetics/131.4.791.

Abstract

We have identified the sua5 locus as a suppressor of an aberrant ATG codon located in the leader region of the cyc1 gene. The sua5-1 allele enhances the iso-1-cytochrome c steady state level in the cyc1-1019 mutant from 2% to approximately 60% of normal (Cyc+) and also confers a marked slow growth (Slg-) phenotype. Suppression is not a consequence of altered transcription initiation at the cyc1 locus. The SUA5 wild-type gene was isolated and sequenced, revealing an open reading frame (ORF) encoding a potential protein of 46,537 Da. SUA5 transcript analyses were consistent with expression of the predicted ORF and Sua5 antisera detected a protein with an apparent molecular mass of 44 kDa. SUA5 was mapped to chromosome VII, immediately adjacent to the PMR1 gene. Hybridization analysis revealed the presence of a related gene on chromosome XII. Neither the SUA5 DNA sequence nor deduced amino acid sequence showed homology to any sequences in the data banks. Disruption of SUA5 conferred the same Cyc+ and Slg- phenotypes as the sua5-1 suppressor, which is the result of a missense mutation, encoding a Ser107----Phe replacement. In addition, sua5 null mutants lack cytochrome a.a3 and fail to grow on lactate or glycerol medium. These results define SUA5 as a new gene encoding a novel protein that is necessary for normal cell growth.

摘要

我们已将sua5基因座鉴定为位于cyc1基因前导区的异常ATG密码子的抑制子。sua5-1等位基因可将cyc1-1019突变体中异-1-细胞色素c的稳态水平从正常(Cyc+)的2%提高到约60%,并赋予明显的生长缓慢(Slg-)表型。抑制并非cyc1基因座转录起始改变的结果。分离并测序了SUA5野生型基因,发现一个开放阅读框(ORF),编码一个潜在的46,537 Da蛋白。SUA5转录本分析与预测的ORF表达一致,Sua5抗血清检测到一个表观分子量为44 kDa的蛋白。SUA5定位于第七条染色体,紧邻PMR1基因。杂交分析显示在第十二条染色体上存在一个相关基因。SUA5的DNA序列和推导的氨基酸序列均与数据库中的任何序列无同源性。SUA5的破坏赋予了与sua5-1抑制子相同的Cyc+和Slg-表型,sua5-1抑制子是一个错义突变的结果,编码Ser107到Phe的替换。此外,sua5缺失突变体缺乏细胞色素a.a3,无法在乳酸或甘油培养基上生长。这些结果将SUA5定义为一个编码对正常细胞生长必需的新型蛋白质的新基因。

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