title: The allosteric HIV-1 integrase inhibitor BI-D affects virion maturation but does not influence packaging of a functional RNA genome.
abstract: The viral integrase (IN) is an essential protein for HIV-1 replication. IN inserts the viral dsDNA into the host chromosome, thereby aided by the cellular co-factor LEDGF/p75. Recently a new class of integrase inhibitors was described: allosteric IN inhibitors (ALLINIs). Although designed to interfere with the IN-LEDGF/p75 interaction to block HIV DNA integration during the early phase of HIV-1 replication, the major impact was surprisingly found on the process of virus maturation during the late phase, causing a reverse transcription defect upon infection of target cells. Virus particles produced in the presence of an ALLINI are misformed with the ribonucleoprotein located outside the virus core. Virus assembly and maturation are highly orchestrated and regulated processes in which several viral proteins and RNA molecules closely interact. It is therefore of interest to study whether ALLINIs have unpredicted pleiotropic effects on these RNA-related processes. We confirm that the ALLINI BI-D inhibits virus replication and that the produced virus is non-infectious. Furthermore, we show that the wild-type level of HIV-1 genomic RNA is packaged in virions and these genomes are in a dimeric state. The tRNAlys3 primer for reverse transcription was properly placed on this genomic RNA and could be extended ex vivo. In addition, the packaged reverse transcriptase enzyme was fully active when extracted from virions. As the RNA and enzyme components for reverse transcription are properly present in virions produced in the presence of BI-D, the inhibition of reverse transcription is likely to reflect the mislocalization of the components in the aberrant virus particle.
authors: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors: Nikki
authors: N
authors: van Bel
authors: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors: Yme
authors: Y
authors: van der Velden
authors: Biodim Mutabilis, Romainville, France.
authors: Damien
authors: D
authors: Bonnard
authors: Biodim Mutabilis, Romainville, France.
authors: Erwann
authors: E
authors: Le Rouzic
authors: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors: Atze T
authors: AT
authors: Das
authors: Biodim Mutabilis, Romainville, France.
authors: Richard
authors: R
authors: Benarous
authors: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors: Ben
authors: B
authors: Berkhout
mesh_terms: D000085:Acetates / Q000737:chemistry / Q000494:pharmacology*; D000494:Allosteric Regulation; D002460:Cell Line; D019281:Dimerization; D016679:Genome, Viral; D057809:HEK293 Cells; D019427:HIV Integrase / Q000737:chemistry* / Q000378:metabolism; D019428:HIV Integrase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D054303:HIV Reverse Transcriptase / Q000737:chemistry / Q000378:metabolism; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D006801:Humans; D011804:Quinolines / Q000737:chemistry / Q000494:pharmacology*; D012343:RNA, Transfer / Q000378:metabolism; D012367:RNA, Viral / Q000302:isolation & purification / Q000378:metabolism*; D019065:Virus Assembly / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000085:Acetates; C578374:BI-D compound; D019428:HIV Integrase Inhibitors; D011804:Quinolines; D012367:RNA, Viral; D012343:RNA, Transfer; D019427:HIV Integrase; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase; C514801:p31 integrase protein, Human immunodeficiency virus 1