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集胞藻PCC 6803株对过氧化氢的差异基因表达及假定的PerR调控子

Differential gene expression in response to hydrogen peroxide and the putative PerR regulon of Synechocystis sp. strain PCC 6803.

作者信息

Li Hong, Singh Abhay K, McIntyre Lauren M, Sherman Louis A

机构信息

Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA.

出版信息

J Bacteriol. 2004 Jun;186(11):3331-45. doi: 10.1128/JB.186.11.3331-3345.2004.

Abstract

We utilized a full genome cDNA microarray to identify the genes that comprise the peroxide stimulon in the cyanobacterium Synechocystis sp. strain PCC 6803. We determined that a gene (slr1738) encoding a protein similar to PerR in Bacillus subtilis was induced by peroxide. We constructed a PerR knockout strain and used it to help identify components of the PerR regulon, and we found that the regulatory properties were consistent with the hypothesis that PerR functions as a repressor. This effort was guided by finding putative PerR boxes in positions upstream of specific genes and by careful statistical analysis. PerR and sll1621 (ahpC), which codes for a peroxiredoxin, share a divergent promoter that is regulated by PerR. We found that isiA, encoding a Chl protein that is induced under low-iron conditions, was strongly induced by a short-term peroxide stress. Other genes that were strongly induced by peroxide included sigD, sigB, and genes encoding peroxiredoxins and Dsb-like proteins that have not been studied yet in this strain. A gene (slr1894) that encoded a protein similar to MrgA in B. subtilis was upregulated by peroxide, and a strain containing an mrgA knockout mutation was highly sensitive to peroxide. A number of genes were downregulated, including key genes in the chlorophyll biosynthesis pathway and numerous regulatory genes, including those encoding histidine kinases. We used PerR mutants and a thioredoxin mutant (TrxA1) to study differential expression in response to peroxide and determined that neither PerR nor TrxA1 is essential for the peroxide protective response.

摘要

我们利用全基因组cDNA微阵列来鉴定组成集胞藻PCC 6803菌株中过氧化物刺激子的基因。我们确定一个编码与枯草芽孢杆菌中PerR相似蛋白的基因(slr1738)可被过氧化物诱导。我们构建了一个PerR基因敲除菌株,并利用它来帮助鉴定PerR调控子的组成部分,我们发现其调控特性与PerR作为阻遏物发挥作用的假说一致。这项工作是通过在特定基因上游位置寻找假定的PerR框以及进行仔细的统计分析来指导的。PerR和编码过氧化物还原酶的sll1621(ahpC)共享一个由PerR调控的不同启动子。我们发现,编码在低铁条件下被诱导的Chl蛋白的isiA在短期过氧化物胁迫下被强烈诱导。其他被过氧化物强烈诱导的基因包括sigD、sigB以及编码过氧化物还原酶和尚未在该菌株中研究过的Dsb样蛋白的基因。一个编码与枯草芽孢杆菌中MrgA相似蛋白的基因(slr1894)被过氧化物上调,并且一个含有mrgA基因敲除突变的菌株对过氧化物高度敏感。许多基因被下调,包括叶绿素生物合成途径中的关键基因以及众多调控基因,包括那些编码组氨酸激酶的基因。我们使用PerR突变体和硫氧还蛋白突变体(TrxA1)来研究对过氧化物的差异表达,并确定PerR和TrxA1对于过氧化物保护反应都不是必需的。

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