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| Status |
Public on Jul 30, 2017 |
| Title |
Reduced protein expression in a virus attenuated by codon deoptimization |
| Organism |
Escherichia coli |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
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 co-expressed downstream genes. We observe no changes in protein abundances of other co-expressed 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.
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| Overall design |
mRNA profiles of T7-infected E. coli from three different strains of T7, across three distinct time points post-infection, with three replicates of each strain-time combination.
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| Contributor(s) |
Jack BR, Boutz DR, Paff ML, Smith BL, Bull JJ, Wilke CO |
| Citation(s) |
28698233 |
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| Submission date |
Mar 13, 2017 |
| Last update date |
May 15, 2019 |
| Contact name |
Claus Wilke |
| Organization name |
University of Texas at Austin
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| Department |
Integrative Biology
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| Lab |
Wilke
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| Street address |
2500 Speedway, A4800
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| City |
Austin |
| State/province |
Texas |
| ZIP/Postal code |
78712 |
| Country |
USA |
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| Platforms (1) |
| GPL21222 |
Illumina NextSeq 500 (Escherichia coli) |
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| Samples (27)
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| Relations |
| BioProject |
PRJNA379044 |
| SRA |
SRP101828 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE96573_RAW.tar |
2.1 Mb |
(http)(custom) |
TAR (of TSV) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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