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首页> 外文期刊>Journal of Virology >Preliminary characterization of coxsackievirus B3 temperature-sensitive mutants.
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Preliminary characterization of coxsackievirus B3 temperature-sensitive mutants.

机译：Coxsackievirus B3温度敏感突变体的初步表征。

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Prototype temperature-sensitive (ts) mutants of a coxsackievirus B3 parent virus capable of replication to similar levels at 34 or 39.5 degrees C were examined for the nature of the temperature-sensitive event restricting replication in HeLa cells at 39.5 degrees C. The ts mutant prototypes represented three different non-overlapping complementation groups. The ts1 mutant (complementation group III) synthesized less than 1% of the infectious genomic RNA synthesized by the coxsackievirus B3 parent virus at 39.5 degrees C and was designated an RNA- mutant. Agarose gel analysis of glyoxal-treated RNA from cells inoculated with ts1 virus revealed that cell RNA synthesis continued in the presence of synthesis of the small amount of viral RNA. This mutant was comparatively ineffective in inducing cell cytopathology and in directing synthesis of viral polypeptides, likely due to the paucity of nascent genomes for translation. The ts5 mutant (complementation group II) directed synthesis of appreciable quantities of both viral genomes (RNA+) and capsid polypeptides; however, assembly of these products into virions occurred at a low frequency, and virions assembled at 39.5 degrees C were highly unstable at that temperature. Shift-down experiments with ts5-inoculated cells showed that capsid precursor materials synthesized at 39.5 degrees C can, after shift to 34 degrees C, be incorporated into ts5 virions. We suggest that the temperature-sensitive defect in this prototype is in the synthesis of one of the capsid polypeptides that cannot renature into the correct configuration required for stability in the capsid at 39.5 degrees C. The ts11 mutant (complementation group I) also synthesized appreciable amounts of viral genomes (RNA+) and viral polypeptides at 39.5 degrees C. Assembly of ts11 virions at 39.5 degrees C occurred at a low frequency, and the stability of these virions at 39.5 degrees C was similar to that of the parent coxsackievirus B3 virions. The temperature-sensitive defect in the ts11 prototype is apparently in assembly. The differences in biochemical properties of the three prototype ts mutants at temperatures above 34 degrees C may ultimately offer insight into the differences in pathogenicity observed in neonatal mice for the three prototype ts mutants.
机译：检查能够在34或39.5摄氏度下以34或39.5℃复制的CoxSackievirus B3父病毒的原型温敏感（TS）突变体用于在39.5℃下限制HeLa细胞中的复制的性质.TS突变体原型代表了三个不同的非重叠互补组。 TS1突变体（互补基团III）合成的是由CoxSackeigirus B3亲本病毒合成的不受感染基因组RNA，在39.5℃下合成，并指定RNA-突变体。从接种TS1病毒接种的细胞的血糖处理RNA的琼脂糖凝胶分析显示，细胞RNA合成在合成少量病毒RNA的情况下继续。该突变体在诱导细胞缩细系系和引导病毒多肽的合成方面相对效果，可能是由于缺乏用于翻译的缺乏基因组的缺乏。 TS5突变体（互补组II）定向了可明显的病毒基因组（RNA +）和衣壳多肽的合成;然而，将这些产物的组装成恶劣的病毒粒子，并且在39.5℃下组装的病毒座在该温度下非常不稳定。用TS5接种细胞的切换实验表明，在将39.5℃下合成的衣壳前体材料可以在将34℃变为34℃后，掺入TS5病毒粒中。我们建议该原型中的温度敏感缺陷在合成衣壳多肽的合成中，其不能重向于39.5℃的衣壳中稳定性所需的正确构型。TS11突变体（互补基团I）也合成了可观病毒基因组（RNA +）和病毒多肽的量为39.5℃。在39.5℃下以低频发生TS11病毒粒子组装，并且这些病毒粒子在39.5℃下的稳定性类似于亲本Coxsackeigirus B3病毒粒子的稳定性。 TS11原型的温度敏感缺陷显然是组装。在34摄氏度高于34摄氏度的温度下三种原型TS突变体的生化特性的差异最终可能对三种原型TS突变体的新生儿小鼠观察到的致病性的差异提供了深度。

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