Structure based modification of Bluetongue virus helicase protein VP6 to produce a viable VP6-truncated BTV.
Structure based modification of Bluetongue virus helicase protein VP6 to produce a viable VP6-truncated BTV.
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DOI：
10.1016/j.bbrc.2014.08.028
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发表时间：
2014-09-05
影响因子：
3.1
通讯作者：
Roy, Polly
中科院分区：
文献类型：
--
作者：
Matsuo, Eiko;Leon, Esther;Matthews, Steve J.;Roy, Polly
对BTV VP 6的NMR分析揭示了两个大的环区域。环（aa 34-130）的丢失不影响蛋白质的整体折叠。BTV复制不需要VP 6的区域（aa 34-92）。VP 6的一个区域（aa 93-130）在病毒复制中起重要作用。蓝舌病毒核心蛋白VP 6是一种依赖ATP水解的RNA解旋酶。然而，尽管进行了大量研究，但VP 6在病毒衣壳内的确切作用及其结构仍不清楚。为了研究VP 6在BTV复制中的需求，我们开始了结构和生物学研究。多核核磁共振谱被分配在组氨酸标记的全长VP 6（329个氨基酸残基）以及几个截短的VP 6变体上。分析表明，一个大的结构域与两个大的环区域，表现出显着的构象交换。其中一个环（氨基酸位置34-130）可以被去除而不影响蛋白质的整体折叠。此外，使用BTV反向遗传学系统，有可能证明VP 6截短的BTV在没有任何辅助VP 6蛋白的情况下在BHK细胞中存活，这表明该环区域的大部分对于BTV复制不是绝对需要的。
NMR analysis on BTV VP6 reveals two large loop regions. The loss of a loop (aa 34–130) does not affect the overall fold of the protein. A region of VP6 (aa 34–92) is not required for BTV replication. A region of VP6 (aa 93–130) plays an essential role in the virus replication. Bluetongue virus core protein VP6 is an ATP hydrolysis dependent RNA helicase. However, despite much study, the precise role of VP6 within the viral capsid and its structure remain unclear. To investigate the requirement of VP6 in BTV replication, we initiated a structural and biological study. Multinuclear nuclear magnetic resonance spectra were assigned on his-tagged full-length VP6 (329 amino acid residues) as well as several truncated VP6 variants. The analysis revealed a large structured domain with two large loop regions that exhibit significant conformational exchange. One of the loops (amino acid position 34–130) could be removed without affecting the overall fold of the protein. Moreover, using a BTV reverse genetics system, it was possible to demonstrate that the VP6-truncated BTV was viable in BHK cells in the absence of any helper VP6 protein, suggesting that a large portion of this loop region is not absolutely required for BTV replication.
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影响因子：
5.4
作者：
Boyce, Mark;Celma, Cristina C. P.;Roy, Polly
通讯作者：
Roy, Polly
影响因子：
64.8
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Grimes, JM;Burroughs, JN;Stuart, DI
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Stuart, DI
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Palmarini M
影响因子：
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作者：
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通讯作者：
Roy, P