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Virion swelling is not required for cotranslational disassembly of cowpea chlorotic mottle virus in vitro.

机译:在体外共翻译cow豆褪绿斑驳病毒不需要病毒颗粒溶胀。

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摘要

The mechanism by which virions of cowpea chlorotic mottle virus (CCMV) disassemble and allow for translation of the virion RNA is not well understood. Previous models have suggested that virion swelling is required to expose the virion RNA for translation in a process referred to as cotranslational disassembly (M. Brisco, R. Hull, and T. M. A. Wilson, Virology 148:210-217, 1986; J. W. Roenhorst, J. W. M. van Lent, and B. J. M. Verduin, Virology 164:91-98, 1988; J. W. Roenhorst, J. M. Verduin, and R. W. Goldbach, Virology 168:138-146, 1989). Previous work in our laboratory has identified point mutations in the CCMV coat protein which result in virions with altered swelling characteristics (J. Fox, F. G. Albert, J. Speir, and M. J. Young, Virology 227:229-233, 1997; J. M. Fox, X. Zhao, J. A. Speir, and M. J. Young, Virology 222:115-122, 1996). The wild-type and mutant CCMV virions were used to correlate virion swelling with the ability of virion RNA to be translated in a cell-free wheat germ extract. Mutant virions unable to swell (cpK42R) are as infectious as wild-type virions in vivo, and the levels of translated encapsidated virion RNA are similar to those of wild-type virions in vitro. Mutant virions capable of swelling but not of disassembling in vitro (cpR26C) are noninfectious and have severely reduced levels of translation of the encapsidated virion RNA in vitro. These studies suggest that virion swelling is not required for the cotranslational disassembly of CCMV. Additionally, the results indicate that there is a pH-dependent structural transition in the virion, other than swelling, that results in the RNA's being exposed for translation in vitro. An alternative model suggesting that cotranslational disassembly of CCMV involves presentation of the virion RNA through the virion fivefold axis is proposed.
机译:cow豆绿斑驳斑驳病毒(CCMV)的病毒粒子分解并允许病毒粒子RNA翻译的机制尚不清楚。先前的模型表明,需要病毒体溶胀以暴露病毒体RNA进行翻译,这一过程称为共翻译反汇编(M. Brisco,R. Hull,and TMA Wilson,Virology 148:210-217,1986; JW Roenhorst,JWM van Lent和BJM Verduin,病毒学,164:91-98,1988; JW Roenhorst,JM Verduin和RW Goldbach,病毒学,168:138-146,1989。我们实验室的先前工作已经确定了CCMV外壳蛋白中的点突变,这些突变导致毒粒的溶胀特性发生了变化(J. Fox,FG Albert,J。Speir和MJ Young,Virology 227:229-233,1997; JM Fox, X.Zhao,JA Speir和MJ Young,Virology 222:115-122,1996)。野生型和突变CCMV病毒体用于将病毒体肿胀与病毒体RNA在无细胞小麦胚芽提取物中翻译的能力相关。无法溶胀的突变病毒体(cpK42R)在体内与野生型病毒体一样具有传染性,并且翻译的衣壳化病毒体RNA的水平与体外野生型病毒体相似。能够溶胀但不能在体外分解的突变病毒体(cpR26C)是非感染性的,并且在体外大大降低了衣壳化病毒体RNA的翻译水平。这些研究表明,CCMV的共翻译拆卸不需要病毒体溶胀。此外,结果表明,在病毒体中,除了溶胀以外,还存在pH依赖性的结构转变,这导致RNA暴露于体外翻译。提出了一个替代模型,该模型表明CCMV的共翻译拆卸涉及通过病毒体五重轴呈递病毒体RNA。

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