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phiEco32-like 噬菌体 7-11 的基因表达调控。

Regulation of Gene Expression of phiEco32-like Bacteriophage 7-11.

机构信息

Institute of Molecular Genetics of National Research Centre «Kurchatov Institute», 123182 Moscow, Russia.

Waksman Institute for Microbiology and Department of Molecular Biology and Biochemistry, Rutgers, State University of New Jersey, Piscataway, NJ 08854, USA.

出版信息

Viruses. 2022 Mar 8;14(3):555. doi: 10.3390/v14030555.

Abstract

serovar Newport bacteriophage 7-11 shares 41 homologous ORFs with phage phiEco32, and both phages encode a protein similar to bacterial RNA polymerase promoter specificity σ subunit. Here, we investigated the temporal pattern of 7-11 gene expression during infection and compared it to the previously determined transcription strategy of phiEco32. Using primer extension and in vitro transcription assays, we identified eight promoters recognized by host RNA polymerase holoenzyme containing 7-11 σ subunit SaPh711_gp47. These promoters are characterized by a bipartite consensus, GTAAtg-(16)-aCTA, and are located upstream of late phage genes. While dissimilar from single-element middle and late promoters of phiEco32 recognized by holoenzymes formed by the phi32_gp36 σ factor, the 7-11 late promoters are located at genome positions similar to those of phiEco32 middle and late promoters. Two early 7-11 promoters are recognized by the RNA polymerase holoenzyme containing the host primary σ factor. Unlike the case of phiEco32, no shut-off of σ-dependent transcription is observed during 7-11 infection and there are no middle promoters. These differences can be explained by the fact that phage 7-11 does not encode a homologue of phi32_gp79, an inhibitor of host and early phage transcription and an activator of transcription by the phi32_gp36-holoenzyme.

摘要

纽波特血清型噬菌体 7-11 与噬菌体 phiEco32 有 41 个同源 ORF,这两种噬菌体都编码一种类似于细菌 RNA 聚合酶启动子特异性 σ 亚基的蛋白质。在这里,我们研究了 7-11 在感染过程中的基因表达时间模式,并将其与之前确定的 phiEco32 转录策略进行了比较。使用引物延伸和体外转录实验,我们鉴定了 8 个被宿主 RNA 聚合酶全酶识别的启动子,该全酶包含 7-11 σ 亚基 SaPh711_gp47。这些启动子的特征是双部分共识 GTAAtg-(16)-aCTA,位于晚期噬菌体基因的上游。虽然与由 phi32_gp36 σ 因子形成的全酶识别的 phiEco32 的单元件中晚期启动子不同,7-11 晚期启动子位于与 phiEco32 中晚期启动子相似的基因组位置。两个早期的 7-11 启动子被包含宿主主要 σ 因子的 RNA 聚合酶全酶识别。与 phiEco32 的情况不同,在 7-11 感染过程中没有观察到 σ 依赖性转录的关闭,也没有中间启动子。这些差异可以用噬菌体 7-11 不编码 phi32_gp79 的同源物来解释,phi32_gp79 是宿主和早期噬菌体转录的抑制剂,也是 phi32_gp36-全酶转录的激活剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8c5a/8948821/c55011ddbb34/viruses-14-00555-g001.jpg

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