The allosteric HIV-1 integrase inhibitor BI-D affects virion maturation but does not influence packaging of a functional RNA genome

PLoS One. 2014 Jul 29;9(7):e103552. doi: 10.1371/journal.pone.0103552. eCollection 2014.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / chemistry
  • Acetates / pharmacology*
  • Allosteric Regulation
  • Cell Line
  • Dimerization
  • Genome, Viral
  • HEK293 Cells
  • HIV Integrase / chemistry*
  • HIV Integrase / metabolism
  • HIV Integrase Inhibitors / chemistry
  • HIV Integrase Inhibitors / pharmacology*
  • HIV Reverse Transcriptase / chemistry
  • HIV Reverse Transcriptase / metabolism
  • HIV-1 / enzymology
  • HIV-1 / genetics
  • HIV-1 / physiology*
  • Humans
  • Quinolines / chemistry
  • Quinolines / pharmacology*
  • RNA, Transfer / metabolism
  • RNA, Viral / isolation & purification
  • RNA, Viral / metabolism*
  • Virus Assembly / drug effects
  • Virus Replication / drug effects*

Substances

  • Acetates
  • BI-D compound
  • HIV Integrase Inhibitors
  • Quinolines
  • RNA, Viral
  • RNA, Transfer
  • HIV Integrase
  • reverse transcriptase, Human immunodeficiency virus 1
  • HIV Reverse Transcriptase
  • p31 integrase protein, Human immunodeficiency virus 1

Grants and funding

This work is supported in part by the Netherlands Organization for Scientific Research (NWO-CW, http://www.nwo.nl/en, Chemical Sciences Division, Top grant 700.59.301 to BB). This work is also supported in part by the European Union Framework Program 7 HIVINNOV (http://www.hivinnov.eu/, Grant 305137). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.