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RNA聚合酶依赖的机制,用于T7噬菌体DNA从病毒体逐步转运到大肠杆菌中。

RNA polymerase-dependent mechanism for the stepwise T7 phage DNA transport from the virion into E. coli.

作者信息

Zavriev S K, Shemyakin M F

出版信息

Nucleic Acids Res. 1982 Mar 11;10(5):1635-52. doi: 10.1093/nar/10.5.1635.

Abstract

The influence of rifampicin, streptolydigin, tetracycline and chloramphenicol on phage DNA transport from the T7 virion into the E. coli cell was studied. It has been found that the DNA transport proceeds in at least three stages. During the initial stage the phage injects into the host cell the left approximately 10 per cent of its DNA molecule. The entrance of the next 50 per cent of 17 DNA molecule is blocked by inhibitors which block transcription but not translation. Moreover, the entrance time of this part of the T7 DNA increases in the case of the T7 mutant D111 (which contains a deletion of the A2 and A3 promoters) and decreases in the case of the D53 mutant (which contains a deletion in the region of the early gene transcription terminator). It would appear, that the second stage of the phage DNA transport is tightly coupled with its transcription and that a mechanical function is carried out by RNA polymerase. The translation inhibitors completely block the entrance of the remaining 40 per cent of the 17 DNA molecule (class III genes) into the host cell. It would appear that some class I and (or) II gene product(s) are obligatory components of the final stage of 17 DNA transport. Some probable consequences of this virus DNA transport model as well as its agreement with the functional structure of T7 chromosome and with T7 development are discussed.

摘要

研究了利福平、链霉溶菌素、四环素和氯霉素对噬菌体DNA从T7病毒粒子转运到大肠杆菌细胞中的影响。已发现DNA转运至少分三个阶段进行。在初始阶段,噬菌体将其DNA分子左侧约10%注入宿主细胞。T7 DNA分子接下来50%的进入被抑制转录但不抑制翻译的抑制剂所阻断。此外,对于T7突变体D111(其A2和A3启动子缺失),这部分T7 DNA的进入时间增加,而对于D53突变体(其早期基因转录终止子区域缺失),进入时间减少。看来,噬菌体DNA转运的第二阶段与其转录紧密耦合,并且RNA聚合酶执行机械功能。翻译抑制剂完全阻断T7 DNA分子剩余40%(III类基因)进入宿主细胞。看来某些I类和(或)II类基因产物是T7 DNA转运最后阶段的必需成分。讨论了这种病毒DNA转运模型的一些可能后果以及它与T7染色体功能结构和T7发育的一致性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fdf5/320555/06e10b050760/nar00374-0218-a.jpg

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