Reduced protein expression in a virus attenuated by codon deoptimization

Benjamin R. Jack, Daniel R. Boutz, Matthew L. Paff, Bartram L. Smith, James J. Bull, Claus O. Wilke

Research output: Contribution to journalArticlepeer-review

Abstract

A general means of viral attenuation involves the extensive recoding of synonymous codons in the viral genome. The mechanistic underpinnings of this approach remain unclear, however. Using quantitative proteomics and RNA sequencing, we explore the molecular basis of attenuation in a strain of bacteriophage T7 whose major capsid gene was engineered to carry 182 suboptimal codons. We do not detect transcriptional effects from recoding. Proteomic observations reveal that translation is halved for the recoded major capsid gene, and a more modest reduction applies to several coexpressed downstream genes. We observe no changes in protein abundances of other coexpressed genes that are encoded upstream. Viral burst size, like capsid protein abundance, is also decreased by half. Together, these observations suggest that, in this virus, reduced translation of an essential polycistronic transcript and diminished virion assembly form the molecular basis of attenuation.

Original languageEnglish (US)
Pages (from-to)2957-2968
Number of pages12
JournalG3: Genes, Genomes, Genetics
Volume7
Issue number9
DOIs
StatePublished - Sep 1 2017

Keywords

  • Bacteriophage T7
  • Codon
  • Deoptimization
  • Recoding
  • Translation
  • Viral attenuation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

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