Suppression of a morphogenic mutant in Rous sarcoma virus capsid protein by a second-site mutation: A cryoelectron tomography study

Carmen Butan, Parvez M. Lokhandwala, John G. Purdy, Giovanni Cardone, Rebecca C. Craven, Alasdair C. Steven

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Retrovirus assembly is driven by polymerization of the Gag polyprotein as nascent virions bud from host cells. Gag is then processed proteolytically, releasing the capsid protein (CA) to assemble de novo inside maturing virions. CA has N-terminal and C-terminal domains (NTDs and CTDs, respectively) whose folds are conserved, although their sequences are divergent except in the 20-residue major homology region (MHR) in the CTD. The MHR is thought to play an important role in assembly, and some mutations affecting it, including the F167Y substitution, are lethal. A temperature-sensitive second-site suppressor mutation in the NTD, A38V, restores infectivity. We have used cryoelectron tomography to investigate the morphotypes of this double mutant. Virions produced at the nonpermissive temperature do not assemble capsids, although Gag is processed normally; moreover, they are more variable in size than the wild type and have fewer glycoprotein spikes. At the permissive temperature, virions are similar in size and spike content as in the wild type and capsid assembly is restored, albeit with altered polymorphisms. The mutation F167Y-A38V (referred to as FY/AV in this paper) produces fewer tubular capsids than wild type and more irregular polyhedra, which tend to be larger than in the wild type, containing ∼30% more CA subunits. It follows that FY/AV CA assembles more efficiently in situ than in the wild type and has a lower critical concentration, reflecting altered nucleation properties. However, its infectivity is lower than that of the wild type, due to a 4-fold-lower budding efficiency. We conclude that the wild-type CA protein sequence represents an evolutionary compromise between competing requirements for optimization of Gag assembly (of the immature virion) and CA assembly (in the maturing virion).

Original languageEnglish (US)
Pages (from-to)6377-6386
Number of pages10
JournalJournal of virology
Volume84
Issue number13
DOIs
StatePublished - Jul 1 2010

Fingerprint

Rous sarcoma virus
tomography
Capsid Proteins
coat proteins
virion
Virion
Tomography
mutation
mutants
Mutation
capsid
Capsid
Temperature
Retroviridae
pathogenicity
inflorescences
Genetic Suppression
gag Gene Products
temperature
lethal genes

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Butan, Carmen ; Lokhandwala, Parvez M. ; Purdy, John G. ; Cardone, Giovanni ; Craven, Rebecca C. ; Steven, Alasdair C. / Suppression of a morphogenic mutant in Rous sarcoma virus capsid protein by a second-site mutation : A cryoelectron tomography study. In: Journal of virology. 2010 ; Vol. 84, No. 13. pp. 6377-6386.
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abstract = "Retrovirus assembly is driven by polymerization of the Gag polyprotein as nascent virions bud from host cells. Gag is then processed proteolytically, releasing the capsid protein (CA) to assemble de novo inside maturing virions. CA has N-terminal and C-terminal domains (NTDs and CTDs, respectively) whose folds are conserved, although their sequences are divergent except in the 20-residue major homology region (MHR) in the CTD. The MHR is thought to play an important role in assembly, and some mutations affecting it, including the F167Y substitution, are lethal. A temperature-sensitive second-site suppressor mutation in the NTD, A38V, restores infectivity. We have used cryoelectron tomography to investigate the morphotypes of this double mutant. Virions produced at the nonpermissive temperature do not assemble capsids, although Gag is processed normally; moreover, they are more variable in size than the wild type and have fewer glycoprotein spikes. At the permissive temperature, virions are similar in size and spike content as in the wild type and capsid assembly is restored, albeit with altered polymorphisms. The mutation F167Y-A38V (referred to as FY/AV in this paper) produces fewer tubular capsids than wild type and more irregular polyhedra, which tend to be larger than in the wild type, containing ∼30{\%} more CA subunits. It follows that FY/AV CA assembles more efficiently in situ than in the wild type and has a lower critical concentration, reflecting altered nucleation properties. However, its infectivity is lower than that of the wild type, due to a 4-fold-lower budding efficiency. We conclude that the wild-type CA protein sequence represents an evolutionary compromise between competing requirements for optimization of Gag assembly (of the immature virion) and CA assembly (in the maturing virion).",
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Suppression of a morphogenic mutant in Rous sarcoma virus capsid protein by a second-site mutation : A cryoelectron tomography study. / Butan, Carmen; Lokhandwala, Parvez M.; Purdy, John G.; Cardone, Giovanni; Craven, Rebecca C.; Steven, Alasdair C.

In: Journal of virology, Vol. 84, No. 13, 01.07.2010, p. 6377-6386.

Research output: Contribution to journalArticle

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AU - Butan, Carmen

AU - Lokhandwala, Parvez M.

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