A SMALL VIRAL-RNA IS REQUIRED FOR INVITRO PACKAGING OF BACTERIOPHAGE-PHI-29 DNA
A SMALL VIRAL-RNA IS REQUIRED FOR INVITRO PACKAGING OF BACTERIOPHAGE-PHI-29 DNA
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DOI：
10.1126/science.3107124
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发表时间：
1987-05-08
期刊：
影响因子：
56.9
通讯作者：
ANDERSON, D
中科院分区：
文献类型：
--
作者：
GUO, PX;ERICKSON, S;ANDERSON, D
枯草芽孢杆菌噬菌体φ-1的小RNA 29在体外病毒DNA包装中具有新的重要作用。在病毒DNA的折叠中对RNA的这种需求为伴随的蛋白质-DNA相互作用提供了新的复杂性维度。RNA是DNA包装机的病毒前体外壳的组成部分，但不是成熟病毒体的组成部分。对涉及这种RNA分子的顺序相互作用的研究可能为小RNA分子的结构和可能的催化作用提供新的见解。φ- 29提取物中的组装和29.核糖核酸酶A或T1处理可强烈抑制体外系统中的DNA包装。然而，噬菌体组装通常发生在核糖核酸酶A的存在下，核糖核酸酶A已经用核糖核酸酶抑制剂处理过。将约120个核苷酸的RNA与φ- 29前体蛋白壳（prohead），并且该颗粒是核糖核酸酶作用的靶标。用核糖核酸酶将DNA从前体中除去，使其对DNA包装失去活性。通过RNA-DNA杂交分析，RNA显示来自非常靠近基因组左端的病毒DNA片段，在体外组装期间，该末端首先包装。
A small RNA of Bacillus subtilis bacteriophage .phi.-29 is shown to have a novel and essential role in viral DNA packaging in vitro. This requirement for RNA in the encapsidation of viral DNA provides a new dimensions of complexity to the attendant protein-DNA interactions. The RNA is a constituent of the viral precursor shell of the DNA-packing machine but is not a component of the mature virion. Studies of the sequential interactions involving this RNA molecule are likely to provide new insight into the structural and possible catalytic roles of small RNA molecules. The .phi.-29 assembly in extracts and .phi.-29 DNA packaging in the defined in vitro system were strongly inhibited by treatment with the ribonucleases A or T1. However, phage assembly occurred normally in the presence of ribonuclease A that had been treated with a ribonuclease inhibitor. An RNA of approximately 120 nucleotides co-purified with the .phi.-29 precursor protein shell (prohead), and this particle was the target of ribonuclease action. Removal of DNA from the prohead by ribonuclease rendered it inactive for DNA packaging. By RNA-DNA hybridization analysis, the RNA was shown to originate from a viral DNA segment very near the left end of the genome, the end packaged first during in vitro assembly.