Martegani E, Brambilla L, Porro D, Ranzi B M, Alberghina L
Dipartimento di Fisiologia e Biochimica Generali, Università di Milano, Italy.
Yeast. 1993 Jun;9(6):575-82. doi: 10.1002/yea.320090603.
Transformed Saccharomyces cerevisiae cells overexpressing the Escherichia coli LacZ gene and the transcriptional activator GAL4, release in the external medium a fraction (from 2 to 10%) of the total beta-galactosidase activity (Porro et al., 1992b). It is known that this abnormal release of a cytoplasmic protein is related to a partial cell lysis of the yeast population, which is likely to be caused by the overexpression of the transcriptional activator GAL4. In the present paper we have characterized the GAL4-induced cell lysis phenomenon. The expression of the GAL4 gene causes morphological modifications and alteration of the cell size distribution. The cell lysis is independent of the expression of the heterologous LacZ gene and occurs in a specific subpopulation of cells (the parent cells) independently of the genealogical age, growth phase conditions and cell cycle progression. Lysis is preceded by a loss of the plasma membrane integrity as indicated by the uptake of ethidium bromide in unfixed cells. Computer analysis of simulated protein distributions indicates that cell lysis takes place in a sizeable aliquot (about 50%) of the parent cells, therefore profoundly altering the age structure of the population.
过表达大肠杆菌LacZ基因和转录激活因子GAL4的转化酿酒酵母细胞,会在外部培养基中释放出总β-半乳糖苷酶活性的一部分(2%至10%)(波罗等人,1992b)。已知这种细胞质蛋白的异常释放与酵母群体的部分细胞裂解有关,这可能是由转录激活因子GAL4的过表达引起的。在本文中,我们对GAL4诱导的细胞裂解现象进行了表征。GAL4基因的表达会导致形态改变和细胞大小分布的变化。细胞裂解与异源LacZ基因的表达无关,且发生在特定的细胞亚群(亲代细胞)中,与系谱年龄、生长阶段条件和细胞周期进程无关。如未固定细胞中溴化乙锭的摄取所示,裂解之前质膜完整性会丧失。对模拟蛋白质分布的计算机分析表明,细胞裂解发生在相当一部分(约50%)的亲代细胞中,因此深刻改变了群体的年龄结构。