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细菌groM基因产物参与噬菌体T7的繁殖。I. 在DNA包装水平上的阻滞。

Involvement of the bacterial groM gene product in bacteriophage T7 reproduction. I. Arrest at the level of DNA packaging.

作者信息

Kuhn A H, Moncany M L, Kellenberger E, Hausmann R

出版信息

J Virol. 1982 Feb;41(2):657-73. doi: 10.1128/JVI.41.2.657-673.1982.

Abstract

The multiplication of bacteriophage T7 is blocked in Escherichia coli M. The genetic determinant of this ability (groM) to inhibit T7 growth was transferred to an E. coli K-12 recipient by means of conjugation. We determined at which precise step T7 maturation is blocked. Phage-directed protein and DNA synthesis as well as degradation of host DNA were not qualitatively affected. Instead of infective phages, only preheads were produced. These, however, were maturable in vitro. The newly synthesized phage DNA accumulated in a concatemeric form and matured from its tetrameric or longer forms (very fast sedimenting DNA) only into its dimeric form (fast-sedimenting DNA) or longer forms. The following step, i.e., the maturation of the dimeric to unit-length DNA, was not observed. Since the concatemeric form of T7 DNA accumulated in spite of the presence of maturable preheads, it is likely that the maturation process was blocked at the level of DNA packaging. As intermediates in the packaging process, we found some prehead-DNA complexes. We interpreted these as true assembly intermediates (or breakdown products thereof), since the attached DNA was still in its concatemeric form. This shows that the very first DNA packaging step, the binding of the progeny DNA to the preheads, was obviously not blocked. Rather, a later step, such as the filling of the preheads with T7 DNA or the stabilization of completely packaged particles (i.e., the final cutting of the concatemers into unit-size length), was inhibited.

摘要

噬菌体T7在大肠杆菌M中的增殖受到阻碍。这种抑制T7生长的能力(groM)的遗传决定因素通过接合转移到了大肠杆菌K-12受体中。我们确定了T7成熟在哪个精确步骤被阻断。噬菌体指导的蛋白质和DNA合成以及宿主DNA的降解在质量上没有受到影响。没有产生感染性噬菌体,只产生了前头部。然而,这些前头部在体外是可成熟的。新合成的噬菌体DNA以串联形式积累,并且仅从其四聚体或更长形式(沉降非常快的DNA)成熟为其二聚体形式(沉降快的DNA)或更长形式。没有观察到下一步,即二聚体DNA成熟为单位长度DNA的过程。尽管存在可成熟的前头部,但T7 DNA仍以串联形式积累,因此成熟过程可能在DNA包装水平上被阻断。作为包装过程的中间体,我们发现了一些前头部-DNA复合物。我们将这些解释为真正的组装中间体(或其分解产物),因为附着的DNA仍处于串联形式。这表明DNA包装的第一步,即子代DNA与前头部的结合,显然没有被阻断。相反,后来的步骤,如用T7 DNA填充前头部或稳定完全包装的颗粒(即最终将串联体切割成单位大小的长度)受到了抑制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e4/256795/45c3059a69aa/jvirol00161-0322-a.jpg

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