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Protein synthesis in long-term stationary-phase cultures of Saccharomyces cerevisiae.

机译:酿酒酵母长期固定期培养中的蛋白质合成。

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We are interested in characterizing the process of entry into and the maintenance of the stationary phase. To identify proteins that are induced during growth to stationary phase, we examined protein synthesis in long-term stationary-phase cultures using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Although the total rate of protein synthesis declined when growth ceased after the postdiauxic phase, the pattern of proteins synthesized remained similar throughout the experimental period (28 days), except at the diauxic shift. At the diauxic shift most proteins detectable by 2D-PAGE undergo a transient reduction in their relative rate of synthesis that ends when cells resume growth during the postdiauxic phase. We conclude from this that the transient repression of protein synthesis at the diauxic shift is not directly associated with stationary-phase arrest. A number of proteins that are synthesized after exponential phase have been identified by 2D-PAGE. These proteins could be divided into three temporal classes depending upon when their synthesis became detectable. One postexponential protein, designated p35, was induced later than all other proteins, and its relative rate of synthesis increased throughout stationary phase. Unlike most postexponential proteins, p35 was not regulated by heat shock or glucose repression. We also observed that a direct correlation between steady-state mRNA accumulation and protein synthesis for another postexponential protein (Ssa3p) or two closely related constitutive proteins (Ssa1p and Ssa2p) did not exist. We conclude from this result that synthesis of proteins in stationary phase is regulated by mechanisms other than the control of steady-state mRNA accumulation.
机译:我们对表征进入和保持固定相的过程感兴趣。为了鉴定在生长到固定相期间诱导的蛋白质,我们使用二维聚丙烯酰胺凝胶电泳(2D-PAGE)检查了长期固定相培养物中的蛋白质合成。尽管在后生期之后生长停止时蛋白质合成的总速率下降了,但合成蛋白的模式在整个实验期间(28天)都保持相似,除了在二生位移时。在双辅助移位中,大多数可通过2D-PAGE检测到的蛋白质的相对合成速率都会发生瞬时降低,这种降低在细胞在双辅助后阶段恢复生长时终止。我们由此得出结论,在双峰位移时蛋白质合成的瞬时抑制与固定相停滞不直接相关。通过2D-PAGE鉴定了指数期后合成的许多蛋白质。根据何时可以检测到它们的合成,可以将这些蛋白质分为三个时间类别。一种称为p35的后指数蛋白比所有其他蛋白晚被诱导,并且其相对合成速率在整个固定相期间均增加。与大多数指数后蛋白不同,p35不受热激或葡萄糖抑制的调节。我们还观察到,稳态mRNA积累与另一后指数蛋白(Ssa3p)或两个紧密相关的组成蛋白(Ssa1p和Ssa2p)的蛋白合成之间没有直接关系。我们从该结果得出结论,固定相蛋白质的合成受稳态mRNA积累控制以外的机制调控。

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