在缺乏完整野生型核衣壳蛋白的情况下包装的莫洛尼鼠白血病病毒基因组 RNA。
Moloney murine leukemia virus genomic RNA packaged in the absence of a full complement of wild type nucleocapsid protein.
机构信息
Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI 48109, USA.
出版信息
Virology. 2012 Sep 1;430(2):100-9. doi: 10.1016/j.virol.2012.05.003. Epub 2012 May 25.
The current model for MLV genomic RNA (gRNA) packaging predicts that of the thousands of Gag proteins in a budding virion, only a small number (≤1%) may be necessary to recruit gRNA. Here, we examined the threshold limits of functional Gag required to package gRNA using wild-type (WT) and packaging deficient mutant nucleocapsid (NC) phenotypically mixed virions. Although gRNA packaging was severely diminished for the NC mutant, the residual encapsidated RNA dimer displayed motility on gels, thermostability, and integrity that was indistinguishable from that of WT. In phenotypically mixed virions, gRNA encapsidation recovered to within approximately two-fold of WT levels when the amount of WT NC was 5-10% of the total. Our results demonstrate that NC's roles in gRNA dimerization and packaging are genetically separable. Additionally, MLV gRNA packaging does not require 100% WT NC, but the amount of functional NC required is greater than the predicted minimum.
目前的 MLV 基因组 RNA（gRNA）包装模型预测，在出芽病毒粒子中的数千个 Gag 蛋白中，只有一小部分（≤1%）可能需要招募 gRNA。在这里，我们使用野生型（WT）和包装缺陷突变体核衣壳（NC）表型混合病毒粒子，检查了包装 gRNA 所需的功能 Gag 的阈值限制。尽管 NC 突变体的 gRNA 包装严重减少，但残留的包裹 RNA 二聚体在凝胶上的迁移率、热稳定性和完整性与 WT 没有区别。在表型混合病毒粒子中，当 WT NC 的量占总 NC 的 5-10%时，gRNA 包裹恢复到 WT 水平的约两倍。我们的结果表明，NC 在 gRNA 二聚化和包装中的作用在遗传上是可分离的。此外，MLV gRNA 包装不需要 100%的 WT NC，但所需的功能 NC 量大于预测的最小值。
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