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大肠杆菌噬菌体H-19B调控区的功能与遗传分析:志贺样毒素和裂解基因的定位表明噬菌体功能在毒素释放中起作用。

Functional and genetic analysis of regulatory regions of coliphage H-19B: location of shiga-like toxin and lysis genes suggest a role for phage functions in toxin release.

作者信息

Neely M N, Friedman D I

机构信息

Department of Microbiology and Immunology, University of Michigan, Ann Arbor 48109-0620, USA.

出版信息

Mol Microbiol. 1998 Jun;28(6):1255-67. doi: 10.1046/j.1365-2958.1998.00890.x.

DOI:10.1046/j.1365-2958.1998.00890.x
PMID:9680214
Abstract

Analysis of the DNA sequence of a 17 kb region of the coli lambdoid phage H-19B genome located the genes encoding shiga-like toxin I (Stx-I) downstream of the gene encoding the analogue of the phage lambda Q transcription activator with its site of action, qut at the associated pR' late promoter, and upstream of the analogues of lambda genes encoding lysis functions. Functional studies, including measurement of the effect of H-19B Q action on levels of Stx expressed from an H-19B prophage, show that the H-19B Q acts as a transcription activator with its associated pR'(qut) by promoting readthrough of transcription terminators. Another toxin-producing phage, 933W, has the identical Q gene and pR'(qut) upstream of the stx-II genes. The H-19B Q also activates Stx-II expression from a 933W prophage. An ORF in H-19B corresponding to the holin lysis genes of other lambdoid phages differs by having only one instead of the usual two closely spaced translation initiation signals that are thought to contribute to the time of lysis. These observations suggest that stx-I expression can be enhanced by transcription from pR' as well as a model for toxin release through cell lysis mediated by action of phage-encoded lysis functions. Functional studies show that open reading frames (ORFs) and sites in H-19B that resemble components of the N transcription antitermination systems controlling early operons of other lambdoid phages similarly promote antitermination. However, this N-like system differs significantly from those of other lambdoid phages.

摘要

对大肠杆菌类λ噬菌体H - 19B基因组17 kb区域的DNA序列分析表明,编码志贺样毒素I(Stx - I)的基因位于编码噬菌体λ Q转录激活因子类似物的基因下游,其作用位点qut位于相关的pR'晚期启动子处,且在编码裂解功能的λ基因类似物的上游。功能研究,包括测量H - 19B Q作用对从H - 19B原噬菌体表达的Stx水平的影响,表明H - 19B Q通过促进转录终止子的通读,作为与其相关的pR'(qut)的转录激活因子发挥作用。另一种产生毒素的噬菌体933W,在stx - II基因上游具有相同的Q基因和pR'(qut)。H - 19B Q也能激活933W原噬菌体中的Stx - II表达。H - 19B中一个与其他类λ噬菌体的穿孔素裂解基因相对应的开放阅读框(ORF)有所不同,它只有一个而非通常的两个紧密间隔的翻译起始信号,人们认为这些信号有助于控制裂解时间。这些观察结果表明,pR'转录可增强stx - I的表达,同时也提出了一种通过噬菌体编码的裂解功能介导的细胞裂解来释放毒素的模型。功能研究表明,H - 19B中类似于控制其他类λ噬菌体早期操纵子的N转录抗终止系统组件的开放阅读框(ORF)和位点同样促进抗终止。然而,这个类似N的系统与其他类λ噬菌体的系统有显著差异。

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