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酿酒酵母酸性磷酸酶突变体的分离与鉴定

Isolation and characterization of acid phosphatase mutants in Saccharomyces cerevisiae.

作者信息

To-E A, Ueda Y, Kakimoto S I, Oshima Y

出版信息

J Bacteriol. 1973 Feb;113(2):727-38. doi: 10.1128/jb.113.2.727-738.1973.

Abstract

Saccharomyces cerevisiae strain H-42 seems to have two kinds of acid phosphatase: one which is constitutive and one which is repressible by inorganic phosphate. The constitutive enzyme was significantly unstable to heat inactivation, and its K(m) of 9.1 x 10(-4)m for p-nitrophenylphosphate was higher than that of the repressible enzyme (2.4 x 10(-4)m). The constitutive and the repressible acid phosphatases are specified by the phoC gene and by the phoB, phoD, or phoE gene, respectively. Results of tetrad analysis suggested that the phoC and phoE genes are linked to the lys2 locus on chromosome II. Since both repressible acid and alkaline phosphatases were affected simultaneously in the phoR, phoD, and phoS mutants, it was concluded that these enzymes were under the same regulatory mechanism or that they shared a common polypeptide. The phoR mutant produced acid phosphatase constitutively, and the phoR mutant allele was recessive to its wild-type counterpart. The phoS mutant showed a phenotype similar to that of a mutant defective in one of the phoB, phoD, or phoE genes. However, the results of genetic analysis of the phoS mutant clearly indicated that the phoS gene is not a structural gene for either of the repressible acid and alkaline phosphatases, but is a kind of regulatory gene. According to the proposed model, the phoS gene controls the expression of the phoR gene, and inorganic phosphate would act primarily as an inducer for the formation of the phoR product which represses phosphatase synthesis.

摘要

酿酒酵母菌株H - 42似乎有两种酸性磷酸酶:一种是组成型的,另一种可被无机磷酸盐抑制。组成型酶对热失活明显不稳定,其对磷酸对硝基苯酯的K(m)为9.1×10(-4)m,高于可抑制酶的K(m)(2.4×10(-4)m)。组成型和可抑制酸性磷酸酶分别由phoC基因和phoB、phoD或phoE基因指定。四分体分析结果表明,phoC和phoE基因与染色体II上的lys2位点连锁。由于在phoR、phoD和phoS突变体中,可抑制的酸性和碱性磷酸酶同时受到影响,因此得出结论,这些酶处于相同的调控机制下,或者它们共享一个共同的多肽。phoR突变体组成型地产生酸性磷酸酶,phoR突变等位基因相对于其野生型对应物是隐性的。phoS突变体表现出与phoB、phoD或phoE基因之一缺陷的突变体相似的表型。然而,phoS突变体的遗传分析结果清楚地表明,phoS基因不是可抑制酸性和碱性磷酸酶中任何一种的结构基因,而是一种调控基因。根据提出的模型,phoS基因控制phoR基因的表达,无机磷酸盐将主要作为phoR产物形成的诱导剂,该产物抑制磷酸酶的合成。

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