RNA IN DNA VIRUS ASSEMBLY

120个核苷酸的噬菌体PHI 29 RNA是结构性的， 病毒前体外壳（prohead）的催化成分转化为 包装18个双链病毒DNA， 在有效的ATP依赖性体外反应中。 述RNA是 探头（连接器）入口顶点的一部分， 与PHI 29 DNA包装蛋白gp 16一起， 作为DNA-gp 3转位所需的ATP酶发挥作用 （PHI 29 DNA在每个5 ′端具有共价结合的蛋白gp 3）。 终点）。 因此，PHI 29 DNA“包装机”包括 连接蛋白（gp 10），RNA，gp 16，也许还有 DNA-gp 3. RNA在序列启动和 DNA-gp 3体外包装的易位事件及其意义 将研究作为ATP酶的一部分的可能功能。 将通过以下方法尝试将RNA定位在连接器中： 电子显微照片和三维图像处理 重建。 PHI 29相关基因和其他基因的前导RNA 将对这些病毒进行分离和测序。 高阶结构 PHI 29前头RNA和对应RNA的研究将通过 使用比较序列分析，足迹法， 化学和酶探针。

A 120 nucleotide bacteriophage PHI29 RNA is structural and catalytic constituent of the viral precursor shell (prohead) into which the 18 kilobase pair double-stranded viral DNA is packaged in an efficient, ATP-dependent, in vitro reaction. The RNA is a part of the portal vertex of the prohead (connector), which together with the PHI29 DNA packaging protein gp16 may function as an ATPase needed for translocation of DNA-gp3 (PHI29 DNA has a covalently bound protein gp3 at each 5' terminus). Thus the PHI29 DNA "packaging machine" includes the connector protein (gp10), RNA, gp16, and perhaps the gp3 of DNA-gp3. The role of the RNA in the sequential initiation and translocation events of in vitro DNA-gp3 packaging and its possible function as part of the ATPase will be investigated. Localization of the RNA in the connector will be attempted by processing of electron micrographs and three-dimensional reconstructions. Prohead RNAs of PHI29 relatives and other phages will be isolated and sequenced. The higher-order structure of PHI29 prohead RNA and counterpart RNAs will be studied by the use of comparative sequence analysis, footprinting, and chemical and enzymatic probes.