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枯草芽孢杆菌噬菌体PBS2诱导的多亚基RNA聚合酶的转录特异性

Transcriptional specificity of a multisubunit RNA polymerase induced by Bacillus subtilis bacteriophage PBS2.

作者信息

Clark S

出版信息

J Virol. 1978 Jan;25(1):224-37. doi: 10.1128/JVI.25.1.224-237.1978.

Abstract

Bacillus subtilis phage PBS2 induced the synthesis of two temporally defined categories of phage-specified transcripts. The transcription of phage "early" genes was induced almost immediately after infection; this RNA synthesis did not require phage protein synthesis. Phage "late" gene transcription, on the other hand, was induced at an intermediate time in the lytic cycle; this RNA synthesis required the production of phage proteins. Both classes of transcription were resistant to the drug rifampin and, therefore, apparently did not require the rifampin-sensitive component of the host RNA polymerase. A rifampin-resistant, DNA-dependent RNA polymerase was purified from bacteria infected with PBS2. The highly purified phage polymerase consisted of five distinct subunits that remained associated during zone centrifugation, isoelectric focusing, and disc gel electrophoresis. The synthesis of each of the five polypeptides was induced at an intermediate time in the phage lytic cycle. As judged by hybridization competition, hybridization to DNA restriction fragments, and RNA-RNA annealing, the phage-induced RNA polymerase preferentially and asymmetrically transcribed PBS2 late genes in vitro. These findings suggest that the PBS2 RNA polymerase controls the expression of phage genes late in the lytic cycle.

摘要

枯草芽孢杆菌噬菌体PBS2诱导合成了两类在时间上有明确界定的噬菌体特异性转录本。噬菌体“早期”基因的转录在感染后几乎立即被诱导;这种RNA合成不需要噬菌体蛋白质合成。另一方面,噬菌体“晚期”基因的转录在裂解周期的中间阶段被诱导;这种RNA合成需要噬菌体蛋白质的产生。这两类转录都对利福平耐药,因此,显然不需要宿主RNA聚合酶中对利福平敏感的成分。从感染了PBS2的细菌中纯化出了一种耐利福平的、依赖DNA的RNA聚合酶。高度纯化的噬菌体聚合酶由五个不同的亚基组成,在区带离心、等电聚焦和圆盘凝胶电泳过程中这些亚基保持结合状态。这五种多肽各自的合成在噬菌体裂解周期的中间阶段被诱导。通过杂交竞争、与DNA限制片段杂交以及RNA-RNA退火判断,噬菌体诱导的RNA聚合酶在体外优先且不对称地转录PBS2晚期基因。这些发现表明,PBS2 RNA聚合酶在裂解周期后期控制噬菌体基因的表达。

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