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 Lopez

Research output: Contribution to journalArticle

37 Citations (Scopus)

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

Fingerprint

Capsid
HIV-1
Capsid Proteins
Antiviral Agents
Virus Uncoating
Growth
Virus Assembly
Ribonucleoproteins
Viral Proteins
Morphogenesis
Fluorescence Microscopy
Electron Microscopy
Salts
Peptides
Temperature
In Vitro Techniques
Proteins

Keywords

  • assembly
  • capsid
  • Gag
  • HIV
  • retrovirus

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Characterization of the In Vitro HIV-1 Capsid Assembly Pathway. / Barklis, Eric; Alfadhli, Ayna; McQuaw, Carolyn; Yalamuri, Suraj; Still, Amelia; Barklis, Robin Lid; Kukull, Ben; Lopez, Claudia.

In: Journal of Molecular Biology, Vol. 387, No. 2, 27.03.2009, p. 376-389.

Research output: Contribution to journalArticle

Barklis, Eric ; Alfadhli, Ayna ; McQuaw, Carolyn ; Yalamuri, Suraj ; Still, Amelia ; Barklis, Robin Lid ; Kukull, Ben ; Lopez, Claudia. / Characterization of the In Vitro HIV-1 Capsid Assembly Pathway. In: Journal of Molecular Biology. 2009 ; Vol. 387, No. 2. pp. 376-389.
@article{81c33d91d646495aa5fdfe71d20c9193,
title = "Characterization of the In Vitro HIV-1 Capsid Assembly Pathway",
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.",
keywords = "assembly, capsid, Gag, HIV, retrovirus",
author = "Eric Barklis and Ayna Alfadhli and Carolyn McQuaw and Suraj Yalamuri and Amelia Still and Barklis, {Robin Lid} and Ben Kukull and Claudia Lopez",
year = "2009",
month = "3",
day = "27",
doi = "10.1016/j.jmb.2009.01.058",
language = "English (US)",
volume = "387",
pages = "376--389",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Characterization of the In Vitro HIV-1 Capsid Assembly Pathway

AU - Barklis, Eric

AU - Alfadhli, Ayna

AU - McQuaw, Carolyn

AU - Yalamuri, Suraj

AU - Still, Amelia

AU - Barklis, Robin Lid

AU - Kukull, Ben

AU - Lopez, Claudia

PY - 2009/3/27

Y1 - 2009/3/27

N2 - 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.

AB - 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.

KW - assembly

KW - capsid

KW - Gag

KW - HIV

KW - retrovirus

UR - http://www.scopus.com/inward/record.url?scp=61649084652&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=61649084652&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2009.01.058

DO - 10.1016/j.jmb.2009.01.058

M3 - Article

C2 - 19356593

AN - SCOPUS:61649084652

VL - 387

SP - 376

EP - 389

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 2

ER -