Folch-Mallol Jorge Luis, Martínez Luz María, Casas Sergio J, Yang Runying, Martínez-Anaya Claudia, López Lorena, Hernández Alejandra, Nieto-Sotelo Jorge
Department of Plant Molecular Biology, Instituto de Biotecnología de la UNAM, 62250 Cuernavaca, Mor., Mexico.
Microbiology (Reading). 2004 Sep;150(Pt 9):2865-2879. doi: 10.1099/mic.0.27144-0.
Living organisms display large differences in stress resistance throughout their life cycles. To study the coordinated regulation of development and stress responses in exponentially growing yeast, mutants that displayed elevated heat-shock resistance at this stage were screened for. Here, two new mutant alleles of CDC25 in Saccharomyces cerevisiae, cdc25-21 and cdc25-22, are described. During exponential growth in glucose at 25 degrees C, these mutants are resistant to heat, oxidative, osmotic and ionic shock, accumulate stress-protein transcripts, show slow growth rates, thick cell walls and glycogen hyperaccumulation and lack cAMP signalling in response to glucose. Genetic and cellular analyses revealed that the stationary-phase phenotypes of cdc25-21 and cdc25-22 mutants are not due to entrance to a G(0) state during exponential growth, but are the result of a prolonged G(1) phase. It was found that, in the W303 background, CDC25 is dispensable for growth in glucose media. However, CDC25 is essential for growth in galactose, in non-fermentable carbon sources and under continuous incubation at 38 degrees C. In conclusion, the function of the catalytic, C-terminal domain of Cdc25p is not only important for fermentative growth, but also for growth in non-fermentable carbon sources and to trigger galactose derepression.
生物体在其整个生命周期中表现出抗逆性的巨大差异。为了研究指数生长的酵母中发育与应激反应的协同调控,我们筛选了在此阶段表现出增强的热休克抗性的突变体。在此,描述了酿酒酵母中CDC25的两个新突变等位基因,cdc25-21和cdc25-22。在25℃的葡萄糖中指数生长期间,这些突变体对热、氧化、渗透和离子冲击具有抗性,积累应激蛋白转录本,生长速率缓慢,细胞壁增厚且糖原过度积累,并且在响应葡萄糖时缺乏cAMP信号传导。遗传和细胞分析表明,cdc25-21和cdc25-22突变体的静止期表型不是由于指数生长期间进入G(0)状态,而是延长的G(1)期的结果。研究发现,在W303背景下,CDC25对于在葡萄糖培养基中的生长是可有可无的。然而,CDC25对于在半乳糖中生长、在非发酵碳源中生长以及在38℃连续培养下是必不可少的。总之,Cdc25p催化性的C末端结构域的功能不仅对发酵生长很重要,而且对在非发酵碳源中的生长以及触发半乳糖去阻遏也很重要。