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Reverse Genetics Mediated Recovery of Infectious Murine Norovirus
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Reduced Protein Expression in a Virus Attenuated by Codon Deoptimization.

Benjamin R Jack1,2,3, Daniel R Boutz3, Matthew L Paff4,2,3

  • 1Department of Integrative Biology, The University of Texas at Austin, Texas 78712 benjamin.r.jack@gmail.com.

G3 (Bethesda, Md.)
|July 13, 2017
PubMed
Summary
This summary is machine-generated.

Extensive synonymous codon recoding in bacteriophage T7 significantly reduces viral translation and virion assembly, leading to viral attenuation. This study clarifies the molecular basis of codon-redefined viral attenuation.

Keywords:
bacteriophage T7codon deoptimizationrecodingtranslationviral attenuation

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetics

Background:

  • Viral attenuation is a strategy to weaken viruses for therapeutic or research purposes.
  • Synonymous codon recoding is a known method for viral attenuation, but its molecular mechanisms are not fully understood.

Purpose of the Study:

  • To investigate the molecular basis of viral attenuation induced by extensive synonymous codon recoding.
  • To elucidate the effects of recoding on viral gene expression and replication.

Main Methods:

  • Quantitative proteomics and RNA sequencing were employed.
  • A strain of bacteriophage T7 was engineered with 182 suboptimal codons in its major capsid gene.

Main Results:

  • Recoding did not significantly alter viral transcription levels.
  • Translation of the recoded major capsid gene was reduced by half, impacting downstream genes.
  • Viral burst size and capsid protein abundance were decreased by 50%.

Conclusions:

  • Reduced translation of essential polycistronic transcripts is a key mechanism of attenuation in this bacteriophage T7 model.
  • Diminished virion assembly contributes to the observed viral attenuation.
  • Synonymous codon recoding impacts viral fitness through translational efficiency and assembly processes.