Li Hong, Singh Abhay K, McIntyre Lauren M, Sherman Louis A
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA.
J Bacteriol. 2004 Jun;186(11):3331-45. doi: 10.1128/JB.186.11.3331-3345.2004.
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.
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。