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鼠伤寒沙门氏菌中编码能量保守型NADH脱氢酶的nuo操纵子的转录调控。

Transcriptional control of the nuo operon which encodes the energy-conserving NADH dehydrogenase of Salmonella typhimurium.

作者信息

Archer C D, Elliott T

机构信息

Department of Microbiology, University of Alabama at Birmingham 35294, USA.

出版信息

J Bacteriol. 1995 May;177(9):2335-42. doi: 10.1128/jb.177.9.2335-2342.1995.

Abstract

The 14 nuo genes encode the subunits of the type I (energy-conserving) NADH dehydrogenase, a key component of the respiratory chain. Salmonella typhimurium, like Escherichia coli, has two enzymes that can oxidize NADH and transfer electrons to ubiquinone, but only the type I enzyme translocates protons across the membrane to generate a proton motive force. Cells with the type I enzyme are energetically more efficient; the role of the type II enzyme (encoded by ndh) is not established, but it may function like a relief valve to allow more rapid NADH recycling. Here, we have investigated transcription of the nuo gene cluster, primarily in S. typhimurium. Studies with polar insertion mutants demonstrate that these genes are arranged as a single, large operon that is expressed from a complex promoter region upstream of nuoA. The DNA sequence of the promoter region was determined, and primer extension analysis of nuo transcripts was used to map four major RNA 5' ends to this region. A set of lac operon fusions to various DNA segments from the nuo promoter region was also constructed. Analysis of these fusions confirmed the presence of at least two nuo promoters. Mutations in the global regulatory genes arcA, oxrA (fnr), crp, cya, and katF were tested for effects on expression of the nuo operon. However, none of the mutations tested had a large effect on expression of type I NADH dehydrogenase.

摘要

14个nuo基因编码I型(能量保守型)NADH脱氢酶的亚基,该酶是呼吸链的关键组成部分。鼠伤寒沙门氏菌与大肠杆菌一样,有两种能够氧化NADH并将电子传递给泛醌的酶,但只有I型酶能将质子跨膜转运以产生质子动力。具有I型酶的细胞在能量方面更高效;II型酶(由ndh编码)的作用尚未明确,但它可能起到安全阀的作用,使NADH能更快地循环利用。在此,我们主要在鼠伤寒沙门氏菌中研究了nuo基因簇的转录。对极性插入突变体的研究表明,这些基因排列成一个单一的大操纵子,从nuoA上游的复杂启动子区域开始表达。确定了启动子区域的DNA序列,并通过对nuo转录本的引物延伸分析将四个主要RNA 5'末端定位到该区域。还构建了一组与nuo启动子区域不同DNA片段的lac操纵子融合体。对这些融合体的分析证实了至少存在两个nuo启动子。测试了全局调控基因arcA、oxrA(fnr)、crp、cya和katF中的突变对nuo操纵子表达的影响。然而,所测试的突变均未对I型NADH脱氢酶的表达产生重大影响。

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