December 13, 1985Science
Expression of the Rous Sarcoma Virus pol Gene by Ribosomal Frameshifting
Population
Cell-free mammalian translation system and Escherichia coli cell-free translation system programmed by a…
Design
Preclinical
Authors
Discussion
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Implication
Supports frameshifting in vitro; leaves open in vivo efficiency, regulation, and relevance to human retroviruses.
Key Points
- To determine whether the primary translation product of the Rous sarcoma virus pol gene arises via ribosomal frameshifting or RNA splicing.
- Synthesized a single RNA species from cloned Rous sarcoma virus DNA using a bacteriophage RNA polymerase.
- Translated the synthetic RNA in cell-free mammalian and Escherichia coli translation systems to assess protein production across intact and deleted gag sequences.
- Cell-free mammalian translation produced both gag and gag-pol fusion proteins at an approximate 20:1 ratio, matching ratios seen in infected cells.
- Frameshifting occurred specifically at the gag-pol junction, remained unaffected by large gag deletions, and showed ten-fold lower efficiency in an Escherichia coli translation system.
Structured PICO
P
Population
Cell-free mammalian translation system and Escherichia coli cell-free translation system programmed by a single RNA species synthesized from cloned viral DNA
O
Outcome
Synthesis of gag protein and gag-pol fusion protein
Demonstrates that the Rous sarcoma virus pol gene is expressed via ribosomal frameshifting to produce a gag-pol fusion protein.
Cite This Study
Jacks et al. (1985) studied this question.