Binding of host factors to stabilized HIV-1 capsid tubes

Anastasia Selyutina, Angel Bulnes-Ramos, Felipe Diaz-Griffero

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

The capsid-binding assay is an in vitro experiment used to determine whether cellular proteins interact with the HIV-1 core. In vitro assembled HIV-1 capsids recapitulate the surface of the HIV-1 core. The assay involves the incubation of in vitro assembled HIV-1 capsid-nucleocapsid (CA-NC) complexes with the protein in question. Subsequently, the mixture is spun through a sucrose cushion using an ultracentrifuge, and the pellet is analyzed for the presence of the protein in question. Although this binding assay is reliable, it is labor intensive and does not contain washing steps. Here we have developed a simpler and faster assay to measure whether a cellular protein is binding to capsid. More importantly, this novel capsid-binding assay contains washing steps. In this assay, we took advantage of the HIV-1 capsid mutant A14C/E45C protein, which is stabilized by disulfide bonds, and is resistant to washing steps. We validated the reliability and specificity of this novel assay by testing the capsid binding ability of TRIMCyp, CPSF6 and MxB with their corresponding controls. Overall, this novel assay provides a reliable and fast methodology to search for novel capsid binders.

Original languageEnglish (US)
Pages (from-to)1-5
Number of pages5
JournalVirology
Volume523
DOIs
Publication statusPublished - Oct 2018

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Keywords

  • Binding
  • CPSF6
  • Capsid
  • Cyp A
  • HIV-1
  • Human TRIM5α-R332P
  • MxB
  • Stabilized tubes
  • TRIMCyp

ASJC Scopus subject areas

  • Virology

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