Forced Complementation between Subgenomic RNAs: Does Human Immunodeficiency Type 1 Virus Reverse Transcription Occur in Viral Core, Cytoplasm, or Early Endosome?

Journal of AIDS and immune research Pub Date : 2015-01-01 Epub Date: 2015-03-02
Weining Han, Yuejin Li, Bernard S Bagaya, Meijuan Tian, Mastooreh Chamanian, Chuanwu Zhu, Jie Shen, Yong Gao
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Abstract

Although the process of reverse transcription is well elucidated, it remains unclear if viral core disruption provides a more cellular or viral milieu for HIV-1 reverse transcription. We have devised a method to require mixing of viral cores or core constituents to produce infectious progeny virus by a bipartite subgenomic RNA (sgRNA) system, in which HIV-1 cplt_R/U5/gag/Δpol and nfl sgRNAs are complementary to each other and when together can complete viral reverse transcription. Only the heterodiploid virus containing both the nfl and cplt_R/U5/gag/Δpol sgRNAs can complete reverse transcription and propagate infectious virus upon de novo infection. Dual exposure of U87.CD4.CXCR4 cells with high titers of the homodimeric nfl and cplt_R/U5/gag/Δpol virus particles did not result in productive virus infection. On the other hand, in early endosomes, the HIV-1 sgRNAs released from viral cores can retain function and complete the reverse transcription and result in productive infection. These findings confirm the assumptions that, in natural infection, HIV-1 cores, and likely other retrovirus cores, remain largely intact and do not mix/fuse in the cytoplasm during the reverse transcription process, and circulating cytoplasmic HIV-1 sgRNA (produced through transfection) could not help the complementary sgRNA in the viral core to complement the reverse transcription process.

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亚基因组rna之间的强制互补:人类免疫缺陷1型病毒逆转录是否发生在病毒核心、细胞质或早期内体?
尽管逆转录的过程已经被很好地阐明,但尚不清楚病毒核心破坏是否为HIV-1逆转录提供了更多的细胞或病毒环境。我们设计了一种方法,需要混合的病毒核心或核心成分产生传染性的子代病毒的双部亚基因组RNA (sgRNA)系统,其中HIV-1 cplt_R/U5/gag/Δpol和nfl sgRNA是互补的,当一起可以完成病毒逆转录。只有同时含有nfl和cplt_R/U5/gag/Δpol sgRNAs的异二倍体病毒才能在新感染时完成逆转录并繁殖感染性病毒。U87.CD4双重暴露。具有高滴度同源二聚体nfl和cplt_R/U5/gag/Δpol病毒颗粒的CXCR4细胞不会导致产生性病毒感染。另一方面,在早期核内体中,从病毒核心释放的HIV-1 sgrna可以保留功能并完成逆转录并导致多产性感染。这些发现证实了这样的假设,即在自然感染中,HIV-1核心以及其他可能的逆转录病毒核心在逆转录过程中基本上保持完整,不会在细胞质中混合/融合,并且循环的细胞质HIV-1 sgRNA(通过转染产生)不能帮助病毒核心中的互补sgRNA补充逆转录过程。
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