title: HIV-1 protease and reverse transcriptase control the architecture of their nucleocapsid partner.
abstract: The HIV-1 nucleocapsid is formed during protease (PR)-directed viral maturation, and is transformed into pre-integration complexes following reverse transcription in the cytoplasm of the infected cell. Here, we report a detailed transmission electron microscopy analysis of the impact of HIV-1 PR and reverse transcriptase (RT) on nucleocapsid plasticity, using in vitro reconstitutions. After binding to nucleic acids, NCp15, a proteolytic intermediate of nucleocapsid protein (NC), was processed at its C-terminus by PR, yielding premature NC (NCp9) followed by mature NC (NCp7), through the consecutive removal of p6 and p1. This allowed NC co-aggregation with its single-stranded nucleic-acid substrate. Examination of these co-aggregates for the ability of RT to catalyse reverse transcription showed an effective synthesis of double-stranded DNA that, remarkably, escaped from the aggregates more efficiently with NCp7 than with NCp9. These data offer a compelling explanation for results from previous virological studies that focused on i) Gag processing leading to nucleocapsid condensation, and ii) the disappearance of NCp7 from the HIV-1 pre-integration complexes. We propose that HIV-1 PR and RT, by controlling the nucleocapsid architecture during the steps of condensation and dismantling, engage in a successive nucleoprotein-remodelling process that spatiotemporally coordinates the pre-integration steps of HIV-1. Finally we suggest that nucleoprotein remodelling mechanisms are common features developed by mobile genetic elements to ensure successful replication.
authors: Laboratoire de Microscopie Moléculaire, UMR 8126: Interactions Moléculaires et Cancer, CNRS, Université Paris Sud-Institut de Cancérologie Gustave Roussy, Villejuif, France; Division de Biochimie, UFR des Sciences de la Vie, Université Pierre et Marie Curie-Paris, France. gilles.mirambeau@free.fr
authors: Gilles
authors: G
authors: Mirambeau
authors: Sébastien
authors: S
authors: Lyonnais
authors: Dominique
authors: D
authors: Coulaud
authors: Laurence
authors: L
authors: Hameau
authors: Sophie
authors: S
authors: Lafosse
authors: Josette
authors: J
authors: Jeusset
authors: Isabelle
authors: I
authors: Borde
authors: Michèle
authors: M
authors: Reboud-Ravaux
authors: Tobias
authors: T
authors: Restle
authors: Robert J
authors: RJ
authors: Gorelick
authors: Eric
authors: E
authors: Le Cam
mesh_terms: D001665:Binding Sites; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D016679:Genome, Viral; D016333:HIV Protease / Q000235:genetics / Q000378:metabolism*; D054303:HIV Reverse Transcriptase / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics; D007700:Kinetics; D008958:Models, Molecular; D019251:Nucleocapsid / Q000737:chemistry / Q000648:ultrastructure*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics
chemical_list: D004277:DNA, Single-Stranded; D004279:DNA, Viral; D012367:RNA, Viral; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase; D016333:HIV Protease; C514803:p16 protease, Human immunodeficiency virus 1
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