Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany.
Mol Microbiol. 2010 Mar;75(6):1495-512. doi: 10.1111/j.1365-2958.2010.07073.x. Epub 2010 Feb 1.
Inheritable bacterial defence systems against phage infection and foreign DNA, termed CRISPR (clustered regularly interspaced short palindromic repeats), consist of cas protein genes and repeat arrays interspaced with sequences originating from invaders. The Cas proteins together with processed small spacer-repeat transcripts (crRNAs) cause degradation of penetrated foreign DNA by unknown mechanisms. Here, we have characterized previously unidentified promoters of the Escherichia coli CRISPR arrays and cas protein genes. Transcription of precursor crRNA is directed by a promoter located within the CRISPR leader. A second promoter, directing cas gene transcription, is located upstream of the genes encoding proteins of the Cascade complex. Furthermore, we demonstrate that the DNA-binding protein H-NS is involved in silencing the CRISPR-cas promoters, resulting in cryptic Cas protein expression. Our results demonstrate an active involvement of H-NS in the induction of the CRISPR-cas system and suggest a potential link between two prokaryotic defence systems against foreign DNA.
可遗传的细菌防御系统可抵御噬菌体感染和外源 DNA,称为 CRISPR(成簇规律间隔短回文重复序列),由 cas 蛋白基因和重复序列组成,重复序列之间穿插着来自入侵物的序列。Cas 蛋白与加工后的短间隔重复转录本(crRNA)一起通过未知机制导致穿透的外源 DNA 降解。在这里,我们已经鉴定了先前未知的大肠杆菌 CRISPR 阵列和 cas 蛋白基因的启动子。前体 crRNA 的转录由位于 CRISPR 先导区的一个启动子指导。第二个启动子,指导 cas 基因转录,位于编码 Cascade 复合物蛋白的基因上游。此外,我们证明 DNA 结合蛋白 H-NS 参与沉默 CRISPR-cas 启动子,导致隐 cas 蛋白表达。我们的研究结果表明 H-NS 积极参与 CRISPR-cas 系统的诱导,并提示了两种针对外源 DNA 的原核防御系统之间的潜在联系。