Characterization of the In Vitro HIV-1 Capsid Assembly Pathway

Eric Barklis, Ayna Alfadhli, Carolyn McQuaw, Suraj Yalamuri, Amelia Still, Robin Lid Barklis, Ben Kukull, Claudia S. López

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

During the morphogenesis of mature human immunodeficiency virus-1 cores, viral capsid proteins assemble conical or tubular shells around viral ribonucleoprotein complexes. This assembly step is mimicked in vitro through reactions in which capsid proteins oligomerize to form long tubes, and this process can be modeled as consisting of a slow nucleation period, followed by a rapid phase of tube growth. We have developed a novel fluorescence microscopy approach to monitor in vitro assembly reactions and have employed it, along with electron microscopy analysis, to characterize the assembly process. Our results indicate that temperature, salt concentration, and pH changes have differential effects on tube nucleation and growth steps. We also demonstrate that assembly can be unidirectional or bidirectional, that growth can be capped, and that proteins can assemble onto the surfaces of tubes, yielding multiwalled or nested structures. Finally, experiments show that a peptide inhibitor of in vitro assembly also can dismantle preexisting tubes, suggesting that such reagents may possess antiviral effects against both viral assembly and uncoating. Our investigations help establish a basis for understanding the mechanism of mature human immunodeficiency virus-1 core assembly and avenues for antiviral inhibition.

Original languageEnglish (US)
Pages (from-to)376-389
Number of pages14
JournalJournal of molecular biology
Volume387
Issue number2
DOIs
StatePublished - Mar 27 2009

Keywords

  • Gag
  • HIV
  • assembly
  • capsid
  • retrovirus

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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