title: Forced Complementation between Subgenomic RNAs: Does Human Immunodeficiency Type 1 Virus Reverse Transcription Occur in Viral Core, Cytoplasm, or Early Endosome?
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.
authors.affiliation: Suzhou Center for Disease Control and Prevention, Suzhou, China.
authors.forename: Weining
authors.initials: W
authors.lastname: Han
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Case Western Reserve University, 10900, Euclid Ave, Cleveland, Ohio 44106, USA.
authors.forename: Yuejin
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, 10900 Euclid Ave, Cleveland, Ohio 44106, USA.
authors.forename: Bernard S
authors.initials: BS
authors.lastname: Bagaya
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Case Western Reserve University, 10900, Euclid Ave, Cleveland, Ohio 44106, USA.
authors.forename: Meijuan
authors.initials: M
authors.lastname: Tian
authors.affiliation: Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, 10900 Euclid Ave, Cleveland, Ohio 44106, USA.
authors.forename: Mastooreh
authors.initials: M
authors.lastname: Chamanian
authors.affiliation: The Fifth People's Hospital, Suzhou, China.
authors.forename: Chuanwu
authors.initials: C
authors.lastname: Zhu
authors.affiliation: Suzhou Center for Disease Control and Prevention, Suzhou, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Shen
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Case Western Reserve University, 10900, Euclid Ave, Cleveland, Ohio 44106, USA; Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, 10900 Euclid Ave, Cleveland, Ohio 44106, USA.
authors.forename: Yong
authors.initials: Y
authors.lastname: Gao
keywords: Human Immunodeficiency Type 1Virus; Reverse transcription; Subgenomic RNAs