Novick P, Osmond B C, Botstein D
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Genetics. 1989 Apr;121(4):659-74. doi: 10.1093/genetics/121.4.659.
Suppressors of a temperature-sensitive mutation (act1-1) in the single actin gene of Saccharomyces cerevisiae were selected that had simultaneously acquired a cold-sensitive growth phenotype. Five genes, called SAC (suppressor of actin) were defined by complementation tests; both suppression and cold-sensitive phenotypes were recessive. Three of the genes (SAC1, SAC2 and SAC3) were subjected to extensive genetic and phenotypic analysis, including molecular cloning. Suppression was found to be allele-specific with respect to actin alleles. The sac mutants, even in ACT1+ genetic backgrounds, displayed phenotypes similar to those of actin mutants, notably aberrant organization of intracellular actin and deposition of chitin at the cell surface. These results are interpreted as being consistent with the idea that the SAC genes encode proteins that interact with actin, presumably as components or controllers of the assembly or stability of the yeast actin cytoskeleton. Two unexpected properties of the SAC1 gene were noted. Disruptions of the gene indicated that its function is essential only at temperatures below about 17 degrees and all sac1 alleles are inviable when combined with act1-2. These properties are interpreted in the context of the evolution of the actin cytoskeleton of yeast.
筛选出了酿酒酵母单肌动蛋白基因中温度敏感突变(act1-1)的抑制子,这些抑制子同时获得了冷敏感生长表型。通过互补试验定义了五个基因,称为SAC(肌动蛋白抑制子);抑制和冷敏感表型均为隐性。其中三个基因(SAC1、SAC2和SAC3)进行了广泛的遗传和表型分析,包括分子克隆。发现抑制作用相对于肌动蛋白等位基因具有等位基因特异性。即使在ACT1+遗传背景下,sac突变体也表现出与肌动蛋白突变体相似的表型,特别是细胞内肌动蛋白的异常组织和几丁质在细胞表面的沉积。这些结果被解释为与以下观点一致,即SAC基因编码与肌动蛋白相互作用的蛋白质,推测其作为酵母肌动蛋白细胞骨架组装或稳定性的组成部分或调控因子。注意到SAC1基因有两个意外特性。该基因的破坏表明其功能仅在约17度以下的温度下才是必需的,并且所有sac1等位基因与act1-2组合时均无法存活。这些特性在酵母肌动蛋白细胞骨架进化的背景下得到了解释。