INVITRO REPLICATION, PACKAGING, AND TRANSCRIPTION OF THE SEGMENTED DOUBLE-STRANDED-RNA GENOME OF BACTERIOPHAGE PHI-6 - STUDIES WITH PROCAPSIDS ASSEMBLED FROM PLASMID-ENCODED PROTEINS
INVITRO REPLICATION, PACKAGING, AND TRANSCRIPTION OF THE SEGMENTED DOUBLE-STRANDED-RNA GENOME OF BACTERIOPHAGE PHI-6 - STUDIES WITH PROCAPSIDS ASSEMBLED FROM PLASMID-ENCODED PROTEINS
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DOI：
10.1128/jb.172.10.5774-5782.1990
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发表时间：
1990-10-01
影响因子：
3.2
通讯作者：
MINDICH, L
中科院分区：
文献类型：
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作者：
GOTTLIEB, P;STRASSMAN, J;MINDICH, L
含脂质噬菌体vphi 6的基因组含有三个双链RNA（dsRNA）片段。我们制备了病毒基因组的cDNA拷贝，并将该材料克隆到质粒中，所述质粒在大肠杆菌和菜豆假单胞菌（Pseudomonasphaseolicola）（vphi 6的天然宿主）中复制。这些质粒指导病毒蛋白质的形成和类似于含有蛋白质P1、P2、P4和P7的病毒前衣壳的结构的组装。我们发现这些颗粒能够吸收病毒单链RNA并合成负链以产生dsRNA结构。一旦dsRNA形成，它就被用作模板，用于在类似于正常转录的反应中产生病毒正链。这些颗粒也能够直接转录外源dsRNA。复制酶反应对v φ 6 RNA是特异性的，对原衣壳是特异性的，并导致产物dsRNA大量掺入颗粒中。这些结果提供了强有力的支持，在该模型中，基因组包装是由预先形成的原衣壳。
The genome of the lipid-containing bacteriophage .vphi.6 contains three segments of double-stranded RNA (dsRNA). We prepared cDNA copies of the viral genome and cloned this material in plasmids that replicate in Eschierichia coli and Pseudomonas phaseolicola, the natural host of .vphi.6. These plasmids direct the formation of viral proteins and the assembly of structures similar to viral procapsids containing proteins P1, P2, P4, and P7. We found that these particles are capable of taking up viral single-stranded RNA and synthesizing the minus strands to produce dsRNA structures. Once the dsRNA is formed, it is then used as a template for the production of viral plus strands in a reaction that resembles normal transcription. The particles were also capable of directly transcribing exogenous dsRNA. The replicase reactions were specific for .vphi.6 RNA, were specific for procapsids, and resulted in substantial incorporation of product dsRNA into particles. These results offer strong support to a model in which genomic packaging is done by preformed procapsids.