Microbiology and Molecular Genetics Program, Department of Microbiology and Immunology, Emory University, Atlanta, Georgia 30329, USA.
Nature. 2013 May 9;497(7448):254-7. doi: 10.1038/nature12048. Epub 2013 Apr 14.
CRISPR/Cas (clustered regularly interspaced palindromic repeats/CRISPR-associated) systems are a bacterial defence against invading foreign nucleic acids derived from bacteriophages or exogenous plasmids. These systems use an array of small CRISPR RNAs (crRNAs) consisting of repetitive sequences flanking unique spacers to recognize their targets, and conserved Cas proteins to mediate target degradation. Recent studies have suggested that these systems may have broader functions in bacterial physiology, and it is unknown if they regulate expression of endogenous genes. Here we demonstrate that the Cas protein Cas9 of Francisella novicida uses a unique, small, CRISPR/Cas-associated RNA (scaRNA) to repress an endogenous transcript encoding a bacterial lipoprotein. As bacterial lipoproteins trigger a proinflammatory innate immune response aimed at combating pathogens, CRISPR/Cas-mediated repression of bacterial lipoprotein expression is critical for F. novicida to dampen this host response and promote virulence. Because Cas9 proteins are highly enriched in pathogenic and commensal bacteria, our work indicates that CRISPR/Cas-mediated gene regulation may broadly contribute to the regulation of endogenous bacterial genes, particularly during the interaction of such bacteria with eukaryotic hosts.
CRISPR/Cas(规律成簇间隔短回文重复/CRISPR 相关)系统是细菌抵御来自噬菌体或外源性质粒的入侵核酸的防御机制。这些系统使用由重复序列侧翼独特间隔区组成的一系列小 CRISPR RNA(crRNA)来识别其靶标,并利用保守的 Cas 蛋白介导靶标降解。最近的研究表明,这些系统可能在细菌生理学中有更广泛的功能,并且尚不清楚它们是否调节内源性基因的表达。在这里,我们证明弗朗西斯氏菌 novicida 的 Cas 蛋白 Cas9 使用一种独特的、小的、CRISPR/Cas 相关 RNA(scaRNA)来抑制编码细菌脂蛋白的内源性转录本。由于细菌脂蛋白引发针对病原体的促炎先天免疫反应,CRISPR/Cas 介导的细菌脂蛋白表达抑制对于 Francisella novicida 抑制这种宿主反应并促进毒力至关重要。由于 Cas9 蛋白在致病和共生细菌中高度富集,我们的工作表明,CRISPR/Cas 介导的基因调控可能广泛有助于内源性细菌基因的调控,特别是在这些细菌与真核宿主相互作用期间。