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大肠杆菌中半乳糖操纵子的同阻遏物。

Isorepressor of the gal regulon in Escherichia coli.

作者信息

Weickert M J, Adhya S

机构信息

Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.

出版信息

J Mol Biol. 1992 Jul 5;226(1):69-83. doi: 10.1016/0022-2836(92)90125-4.

Abstract

Inducible overexpression of the Escherichia coli gal operon in the absence of the Gal repressor is known as ultrainduction. The requirement of induction can be eliminated by mutation of a new locus, galS, resulting in constitutive and ultrainduced levels of gal expression. Characterization of the galS gene and its product has revealed an isorepressor of the gal regulon. The Gal isorepressor is a protein of 346 amino acid residues whose amino acid sequence and cellular function, as described here, are very similar to that of Gal repressor, encoded by the galR gene. Transcription from different promoters of the gal regulon, galP1, galP2 and mglP, was examined by primer extension and reverse transcription of mRNA isolated from strains containing mutations in galR and/or galS. In strains containing a galS mutation, overexpression of gal message occurred only in the presence of inducer, while mgl message was constitutively derepressed. The galS mutation also constitutively derepressed an mglA::lacZ fusion, demonstrating that GalS is the mgl repressor. A potential operator site in the mgl promoter was identified at a position analogous to OE in gal. Thus, the gal and mgl operons constitute a regulon. Crosstalk, temporal action, induction spectrum or heteromer formation between repressor and isorepressor may help co-ordinate high affinity galactose transport and galactose utilization.

摘要

在没有半乳糖阻遏物的情况下,大肠杆菌半乳糖操纵子的可诱导过表达被称为超诱导。通过一个新位点galS的突变可以消除诱导的需求,从而导致半乳糖表达的组成型和超诱导水平。galS基因及其产物的特征揭示了半乳糖调节子的一种同型阻遏物。半乳糖同型阻遏物是一种由346个氨基酸残基组成的蛋白质,其氨基酸序列和细胞功能(如本文所述)与由galR基因编码的半乳糖阻遏物非常相似。通过引物延伸和对从含有galR和/或galS突变的菌株中分离的mRNA进行逆转录,检测了半乳糖调节子不同启动子galP1、galP2和mglP的转录情况。在含有galS突变的菌株中,半乳糖信息的过表达仅在诱导剂存在时发生,而mgl信息则组成型去阻遏。galS突变也组成型地去阻遏了mglA::lacZ融合体,表明GalS是mgl阻遏物。在mgl启动子中一个类似于gal中OE的位置鉴定出一个潜在的操纵子位点。因此,半乳糖和mgl操纵子构成一个调节子。阻遏物和同型阻遏物之间的串扰、时间作用、诱导谱或异源二聚体形成可能有助于协调高亲和力的半乳糖转运和半乳糖利用。

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