Laboratory of Cellular Biophysics, The Rockefeller University, New York, New York.
Department of Cell Biology, Yale School of Medicine, New Haven, Connecticut.
Traffic. 2018 Jan;19(1):36-43. doi: 10.1111/tra.12536. Epub 2017 Dec 1.
Retroviruses incorporate specific host cell RNAs into virions. In particular, the host noncoding 7SL RNA is highly abundant in all examined retroviruses compared with its cellular levels or relative to common mRNAs such as actin. Using live cell imaging techniques, we have determined that the 7SL RNA does not arrive with the HIV-1 RNA genome. Instead, it is recruited contemporaneously with assembly of the protein HIV-1 Gag at the plasma membrane. Further, we demonstrate that complexes of 7SL RNA and Gag can be immunoprecipitated from both cytosolic and plasma membrane fractions. This indicates that 7SL RNAs likely interact with Gag prior to high-order Gag multimerization at the plasma membrane. Thus, the interactions between Gag and the host RNA 7SL occur independent of the interactions between Gag and the host endosomal sorting complex required for transport (ESCRT) proteins, which are recruited temporarily at late stages of assembly. The interactions of 7SL and Gag are also independent of interactions of Gag and the HIV-1 genome which are seen on the plasma membrane prior to assembly of Gag.
逆转录病毒将特定的宿主细胞 RNA 整合到病毒粒子中。特别是,与细胞水平或与肌动蛋白等常见 mRNA 相比,所有检查过的逆转录病毒中宿主非编码 7SL RNA 的丰度都非常高。使用活细胞成像技术,我们已经确定 7SL RNA 不会与 HIV-1 RNA 基因组一起到达。相反,它是与 HIV-1 Gag 蛋白在质膜上的组装同时被募集的。此外,我们证明来自细胞质和质膜部分的 7SL RNA 和 Gag 复合物可以被免疫沉淀。这表明 7SL RNA 可能在高阶 Gag 多聚体在质膜上形成之前与 Gag 相互作用。因此,Gag 与宿主 RNA 7SL 之间的相互作用发生在 Gag 与内体分选复合物(ESCRT)蛋白之间的相互作用之前,ESCRT 蛋白在组装的晚期被临时募集。7SL 和 Gag 之间的相互作用也独立于 Gag 与 HIV-1 基因组之间的相互作用,在 Gag 组装之前可以在质膜上看到这种相互作用。