Department of Biology, Stanford University, Stanford, California, USA.
J Bacteriol. 2012 Sep;194(18):4983-94. doi: 10.1128/JB.00449-12. Epub 2012 Jul 6.
Sinorhizobium meliloti can live as a soil saprophyte and can engage in a nitrogen-fixing symbiosis with plant roots. To succeed in such diverse environments, the bacteria must continually adjust gene expression. Transcriptional plasticity in eubacteria is often mediated by alternative sigma (σ) factors interacting with core RNA polymerase. The S. meliloti genome encodes 14 of these alternative σ factors, including two putative RpoH ("heat shock") σ factors. We used custom Affymetrix symbiosis chips to characterize the global transcriptional response of S. meliloti rpoH1, rpoH2, and rpoH1 rpoH2 mutants during heat shock and stationary-phase growth. Under these conditions, expression of over 300 genes is dependent on rpoH1 and rpoH2. We mapped transcript start sites of 69 rpoH-dependent genes using 5' RACE (5' rapid amplification of cDNA ends), which allowed us to determine putative RpoH1-dependent, RpoH2-dependent, and dual-promoter (RpoH1- and RpoH2-dependent) consensus sequences that were each used to search the genome for other potential direct targets of RpoH. The inferred S. meliloti RpoH promoter consensus sequences share features of Escherichia coli RpoH promoters but lack extended -10 motifs.
根瘤农杆菌能够作为土壤腐生物生存,并与植物根系进行固氮共生。为了在如此多样化的环境中取得成功,细菌必须不断调整基因表达。真细菌中的转录可塑性通常由与核心 RNA 聚合酶相互作用的替代 sigma(σ)因子介导。S. meliloti 基因组编码了 14 种这样的替代 σ 因子,其中包括两个推定的 RpoH(“热休克”)σ因子。我们使用定制的 Affymetrix 共生芯片来描述 S. meliloti rpoH1、rpoH2 和 rpoH1 rpoH2 突变体在热休克和静止期生长过程中的全局转录响应。在这些条件下,超过 300 个基因的表达依赖于 rpoH1 和 rpoH2。我们使用 5' RACE(5' cDNA 末端快速扩增)来绘制 69 个 rpoH 依赖性基因的转录起始位点,这使我们能够确定推定的 RpoH1 依赖性、RpoH2 依赖性和双启动子(RpoH1 和 RpoH2 依赖性)的共识序列,每个序列都用于在基因组中搜索其他潜在的 RpoH 直接靶标。推断的 S. meliloti RpoH 启动子共识序列具有大肠杆菌 RpoH 启动子的特征,但缺乏扩展的-10 基序。