Department of Microbiology and Immunology, Brody School of Medicine, East Carolina University, Greenville, North Carolina, USA.
J Bacteriol. 2012 Jan;194(1):3-14. doi: 10.1128/JB.05623-11. Epub 2011 Oct 21.
The type IV secretion system encoded by the virB operon is required for full virulence of Brucella sp., and the present study links the RNA chaperone Hfq to wild-type expression of virB in Brucella abortus 2308. Studies employing virB-lacZ fusions, quantitative reverse transcription-PCR, and immunoblot analysis showed that both transcription and translation of virB are decreased in an isogenic hfq mutant compared to those in the parental strain. These results led to the hypothesis that Hfq regulation of virB is mediated through an intermediate transcriptional regulator. Subsequent experiments determined that expression of the gene encoding the putative Brucella quorum-sensing regulator BabR (also known as BlxR), a known virB regulator, is also controlled by Hfq at the posttranscriptional level, and a cis-acting element in the 5' untranslated region of the babR transcript responsible for this regulation was identified. Consistent with its role as a virB regulator, recombinant Brucella BabR binds to the virB promoter region in electrophoretic mobility shift assays. However, experiments employing a babR mutant strain determined that BabR is a repressor, not an activator, of virB transcription. These findings suggest that Hfq regulates virB expression through both BabR-dependent and BabR-independent pathways.
由 virB 操纵子编码的 IV 型分泌系统是布鲁氏菌属完全毒力所必需的,本研究将 RNA 伴侣蛋白 Hfq 与布鲁氏菌 abortus 2308 中野生型 virB 的表达联系起来。采用 virB-lacZ 融合、定量逆转录-PCR 和免疫印迹分析的研究表明,与亲本菌株相比,hfq 突变体中 virB 的转录和翻译均降低。这些结果导致假设 Hfq 对 virB 的调节是通过中间转录调节剂介导的。随后的实验确定,编码假定的布鲁氏菌群体感应调节因子 BabR(也称为 BlxR)的基因表达也受到 Hfq 的转录后调控,并且鉴定了 babR 转录本 5'非翻译区中负责这种调控的顺式作用元件。与它作为 virB 调节剂的作用一致,重组布鲁氏菌 BabR 在电泳迁移率变动分析中结合到 virB 启动子区域。然而,采用 babR 突变株的实验确定 BabR 是 virB 转录的抑制剂,而不是激活剂。这些发现表明,Hfq 通过依赖 BabR 和不依赖 BabR 的途径调节 virB 的表达。