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营养物质诱导的酿酒酵母营养饥饿细胞中海藻糖酶的激活:环磷酸腺苷不充当第二信使。

Nutrient-induced activation of trehalase in nutrient-starved cells of the yeast Saccharomyces cerevisiae: cAMP is not involved as second messenger.

作者信息

Hirimburegama K, Durnez P, Keleman J, Oris E, Vergauwen R, Mergelsberg H, Thevelein J M

机构信息

Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit te Leuven, Flanders, Belgium.

出版信息

J Gen Microbiol. 1992 Oct;138(10):2035-43. doi: 10.1099/00221287-138-10-2035.

Abstract

Starvation of Saccharomyces cerevisiae cells for specific nutrients such as nitrogen, phosphate or sulphate causes arrest in the G1 phase of the cell cycle at a specific point called 'start'. Re-addition of different nitrogen sources, phosphate or sulphate to such starved cells causes activation of trehalase within a few minutes. Nitrogen-source- and sulphate-induced activation of trehalase were not associated with any change in the cAMP level, but in the case of phosphate there was a small transient increase. When nitrogen-source-activated trehalase was isolated by immuno-affinity chromatography from crude extracts, the purified enzyme showed the same activity profile as in the original crude extracts, indicating that post-translational modification is responsible for the activation. In the yeast mutants cdc25-5 and cdc35-10, which are temperature sensitive for cAMP synthesis, incubation at the restrictive temperature lowered but did not prevent nitrogen-, phosphate- or sulphate-induced activation of trehalase. Since under these conditions the cAMP level in the cells is very low, it is unlikely that cAMP acts as a second messenger in this nutrient-induced effect. Nitrogen-source-induced activation of trehalase requires the presence of glucose at a concentration similar to that able to stimulate the RAS-adenylate cyclase pathway. This indicates that the same glucose-sensing system might be involved in both phenomena. Nitrogen-starved cells fractionated according to cell size all showed nitrogen-source-induced activation of trehalase to the same extent, indicating that the nitrogen-induced signalling pathway involved is not dependent on the well-known cell size requirement for progression over the start point of the cell cycle.

摘要

将酿酒酵母细胞置于缺乏特定营养物质(如氮、磷或硫)的环境中,会导致细胞周期在一个特定的点(称为“起始点”)停滞于G1期。向这些饥饿的细胞重新添加不同的氮源、磷或硫,会在几分钟内激活海藻糖酶。氮源和硫诱导的海藻糖酶激活与cAMP水平的任何变化均无关,但在磷的情况下,cAMP水平会有一个短暂的小幅升高。当通过免疫亲和层析从粗提物中分离出氮源激活的海藻糖酶时,纯化后的酶表现出与原始粗提物相同的活性谱,这表明翻译后修饰是激活的原因。在对cAMP合成具有温度敏感性的酵母突变体cdc25 - 5和cdc35 - 10中,在限制温度下孵育会降低但不会阻止氮、磷或硫诱导的海藻糖酶激活。由于在这些条件下细胞内的cAMP水平非常低,因此cAMP不太可能在这种营养诱导效应中作为第二信使起作用。氮源诱导的海藻糖酶激活需要存在浓度与能够刺激RAS - 腺苷酸环化酶途径的浓度相似的葡萄糖。这表明相同的葡萄糖感应系统可能参与了这两种现象。根据细胞大小对氮饥饿细胞进行分级分离,所有细胞均显示出相同程度的氮源诱导的海藻糖酶激活,这表明所涉及的氮诱导信号通路不依赖于细胞周期越过起始点所熟知的细胞大小要求。

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