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大肠杆菌钼酸盐转运操纵子modABCD响应钼酸盐可利用性的调控

Regulation of the molybdate transport operon, modABCD, of Escherichia coli in response to molybdate availability.

作者信息

Rech S, Deppenmeier U, Gunsalus R P

机构信息

Department of Microbiology and Molecular Genetics, University of California, Los Angeles 90024.

出版信息

J Bacteriol. 1995 Feb;177(4):1023-9. doi: 10.1128/jb.177.4.1023-1029.1995.

Abstract

The mod (chlD) locus at 17 min on the Escherichia coli chromosome encodes a high-affinity molybdate uptake system. To further investigate the structure and regulation of these genes, the DNA region upstream of the previously identified modBC (chlJD) genes was cloned and sequenced. A single open reading frame, designated modA, was identified and appears to encode a periplasmic binding protein for the molybdate uptake system. To determine how the mod genes are regulated in response to molybdate, nitrate, and oxygen, we constructed a series of mod-lacZ operon fusions to the upstream region and introduced them in single copy onto the E. coli chromosome. Whereas molybdate limitation resulted in elevated mod-lacZ expression, neither oxygen nor nitrate had any significant effect on gene expression. A regulatory motif, CATAA, located at the modA promoter was identified and shown to be required for molybdate-dependent control of the modABCD operon. Mutations within this sequence resulted in nearly complete derepression of gene expression and suggest that transcription of the operon is mediated by a molybdenum-responsive regulatory protein.

摘要

大肠杆菌染色体上17分钟处的mod(chlD)位点编码一种高亲和力的钼酸盐摄取系统。为了进一步研究这些基因的结构和调控,对先前鉴定的modBC(chlJD)基因上游的DNA区域进行了克隆和测序。鉴定出一个单一的开放阅读框,命名为modA,它似乎编码钼酸盐摄取系统的周质结合蛋白。为了确定mod基因如何响应钼酸盐、硝酸盐和氧气进行调控,我们构建了一系列mod-lacZ操纵子融合体到上游区域,并将它们单拷贝导入大肠杆菌染色体。虽然钼酸盐限制导致mod-lacZ表达升高,但氧气和硝酸盐对基因表达均无显著影响。在modA启动子处鉴定出一个调控基序CATAA,它被证明是modABCD操纵子依赖钼酸盐调控所必需的。该序列内的突变导致基因表达几乎完全去阻遏,这表明操纵子的转录由一种钼响应调节蛋白介导。

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本文引用的文献

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Molybdenum uptake in Escherichia coli K12.大肠杆菌K12对钼的摄取
J Gen Microbiol. 1993 Aug;139(8):1869-75. doi: 10.1099/00221287-139-8-1869.
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Molecular genetic analysis of the moa operon of Escherichia coli K-12 required for molybdenum cofactor biosynthesis.
Mol Microbiol. 1993 Jun;8(6):1071-81. doi: 10.1111/j.1365-2958.1993.tb01652.x.
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Sequence of the malK gene in E.coli K12.大肠杆菌K12中malK基因的序列。
Nucleic Acids Res. 1982 Nov 25;10(22):7449-58. doi: 10.1093/nar/10.22.7449.
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Cloning and nucleotide sequence of the chlD locus.chlD基因座的克隆与核苷酸序列
J Bacteriol. 1987 May;169(5):1911-6. doi: 10.1128/jb.169.5.1911-1916.1987.

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