Analysis of Rous sarcoma virus capsid protein variants assembled on lipid monolayers

Keith Mayo, Marcy L. Vana, Jason McDermott, Doug Huseby, Jonathan Leis, Eric Barklis

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

During assembly and morphogenesis of Rous sarcoma virus (RSV), proteolytic processing of the structural precursor (Pr76Gag) protein generates three capsid (CA) protein variants, CA476, CA479, and CA488. The proteins share identical N-terminal domains (NTDs), but are truncated at residues corresponding to gag codons 476, 479, and 488 in their CA C-terminal domains (CTDs). To characterize oligomeric forms of the RSV CA variants, we examined 2D crystals of the capsid proteins, assembled on lipid monolayers. Using electron microscopy and image analysis approaches, the CA proteins were observed to organize in hexagonal (p6) arrangements, where rings of membrane-proximal NTD hexamers were spaced at 95 Å intervals. Differences between the oligomeric structures of the CA variants were most evident in membrane-distal regions, where apparent CTDs interconnect hexamer rings. In this region, CA488 connections were observed readily, while CA476 and CA479 contacts were resolved poorly, suggesting that in vivo processing of CA488 to the shorter forms may permit virions to adopt a dissembly-competent conformation. In addition to crystalline arrays, the CA479 and CA488 proteins formed small spherical particles with diameters of 165-175 Å. The spheres appear to be arranged from hexamer or hexamer plus pentamer ring subunits that are related to the 2D crystal forms. Our results implicate RSV CA hexamer rings as basic elements in the assembly of RSV virus cores.

Original languageEnglish (US)
Pages (from-to)667-678
Number of pages12
JournalJournal of molecular biology
Volume316
Issue number3
DOIs
StatePublished - Jan 1 2002

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Keywords

  • Capsid
  • Electron microscopy
  • Gag
  • Retrovirus
  • Rous sarcoma virus

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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