φX174 procapsid assembly: Effects of an inhibitory external scaffolding protein and resistant coat proteins in vitro

James E. Cherwa, Joshua Tyson, Gregory J. Bedwell, Dewey Brooke, Ashton G. Edwards, Terje Dokland, Peter E. Prevelige, Bentley A Fane

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

During φX174 morphogenesis, 240 copies of the external scaffolding protein D organize 12 pentameric assembly intermediates into procapsids, a reaction reconstituted in vitro. In previous studies, φX174 strains resistant to exogenously expressed dominant lethal D genes were experimentally evolved. Resistance was achieved by the stepwise acquisition of coat protein mutations. Once resistance was established, a stimulatory D protein mutation that greatly increased strain fitness arose. In this study, in vitro biophysical and biochemical methods were utilized to elucidate the mechanistic details and evolutionary trade-offs created by the resistance mutations. The kinetics of procapsid formation was analyzed in vitro using wild-type, inhibitory, and experimentally evolved coat and scaffolding proteins. Our data suggest that viral fitness is correlated with in vitro assembly kinetics and demonstrate that in vivo experimental evolution can be analyzed within an in vitro biophysical context.

Original languageEnglish (US)
Article numbere01878-16
JournalJournal of Virology
Volume91
Issue number1
DOIs
StatePublished - 2017

Fingerprint

scaffolding proteins
Capsid Proteins
coat proteins
mutation
kinetics
Proteins
Mutation
lethal genes
in vitro studies
morphogenesis
Lethal Genes
Dominant Genes
Morphogenesis
In Vitro Techniques
genes
proteins
methodology

Keywords

  • Bacteriophage φX174
  • Microviridae
  • Microvirus
  • Scaffolding protein
  • Virus assembly

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

φX174 procapsid assembly : Effects of an inhibitory external scaffolding protein and resistant coat proteins in vitro. / Cherwa, James E.; Tyson, Joshua; Bedwell, Gregory J.; Brooke, Dewey; Edwards, Ashton G.; Dokland, Terje; Prevelige, Peter E.; Fane, Bentley A.

In: Journal of Virology, Vol. 91, No. 1, e01878-16, 2017.

Research output: Contribution to journalArticle

Cherwa, JE, Tyson, J, Bedwell, GJ, Brooke, D, Edwards, AG, Dokland, T, Prevelige, PE & Fane, BA 2017, 'φX174 procapsid assembly: Effects of an inhibitory external scaffolding protein and resistant coat proteins in vitro', Journal of Virology, vol. 91, no. 1, e01878-16. https://doi.org/10.1128/JVI.01878-16
Cherwa, James E. ; Tyson, Joshua ; Bedwell, Gregory J. ; Brooke, Dewey ; Edwards, Ashton G. ; Dokland, Terje ; Prevelige, Peter E. ; Fane, Bentley A. / φX174 procapsid assembly : Effects of an inhibitory external scaffolding protein and resistant coat proteins in vitro. In: Journal of Virology. 2017 ; Vol. 91, No. 1.
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AU - Prevelige, Peter E.

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