Rebinding of transcriptase components (L and NS proteins) to the nucleocapsid template of vesicular stomatitis virus
Rebinding of transcriptase components (L and NS proteins) to the nucleocapsid template of vesicular stomatitis virus
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DOI：
10.1128/jvi.27.3.560-567.1978
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发表时间：
1978-09
影响因子：
5.4
通讯作者：
M. Mellon;S. Emerson
中科院分区：
文献类型：
--
作者：
M. Mellon;S. Emerson
用 Triton X-100 和高盐缓冲液溶解水泡性口炎病毒粒子（新泽西血清型）的 L 和 NS 蛋白，并在类似于体外重建转录酶活性的条件下与纯化的核衣壳重组。在甘油梯度上将核衣壳结合的 L 和 NS 蛋白与未结合的蛋白分离。 L和NS蛋白的重新结合模拟了体内结合，在饱和状态下，L和NS分子与N分子的比率与在完整病毒体中观察到的大致相同。 L 和 NS 蛋白被分离并独立地和组合地添加回模板。在 L 蛋白不存在的情况下，纯化的 NS 蛋白与模板结合。然而，L 蛋白结合似乎取决于 NS 蛋白的存在。转录所需的 Mg2+ 和核苷酸的存在对于 L 和 NS 蛋白的重新结合不是必需的。
The L and NS proteins of vesicular stomatitis virions (New Jersey serotype) were solubilized with Triton X-100 and high-salt buffer and recombined with purified nucleocapsids under conditions similar to those used to reconstitute transcriptase activity in vitro. The nucleocapsid-bound L and NS proteins were separated from unbound proteins on a glycerol gradient. The rebinding of L and NS proteins mimics the in vivo binding in that at saturation the ratio of L and NS molecules to N molecules is approximately the same as observed in the intact virion. L and NS proteins were separated and added back independently and in combination to the template. The purified NS protein bound to the template in the absence of L protein. However, the L protein binding appeared to depend on the presence of NS protein. The presence of Mg2+ and nucleotides, which is required for transcription, was not necessary for the rebinding of L and NS proteins.