Reverse transcriptase of Moloney murine leukemia virus binds to eukaryotic release factor 1 to modulate suppression of translational termination.
The pol (for polymerase) gene of the murine leukemia viruses (MuLVs) is expressed in the form of a large Gag-Pol precursor protein by the suppression of translational termination, or enhanced readthrough, of a UAG stop codon at the end of gag. A search for cellular proteins that interact with the reverse transcriptase of Moloney MuLV resulted in the identification of eRF1, the eukaryotic translation release factor 1. The proteins bound strongly in vitro, and the overexpression of eRF1 resulted in the RT-dependent incorporation of the protein into assembling virion particles. The overexpression of RT in trans enhanced the translational readthrough of a reporter construct containing the Gag-Pol boundary region. Noninteracting mutants of RT failed to synthesize adequate levels of Gag-Pol and could not replicate. These results suggest that RT enhances suppression of termination and that the interaction of RT with eRF1 is required for an appropriate level of translational readthrough.
Orlova
Marianna
M
Department of Biochemistry and Molecular Biophysics, Integrated Program in Cell and Molecular Biology, Howard Hughes Medical Institute, Columbia University College of Physicians and Surgeons, New York, NY 10032, USA.
Yueh
Andrew
A
Leung
Juliana
J
Goff
Stephen P
SP
D000818:Animals; D001667:Binding, Competitive; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism; D015685:Fusion Proteins, gag-pol / Q000096:biosynthesis / Q000378:metabolism; D051379:Mice; D008979:Moloney murine leukemia virus / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D009154:Mutation; D010443:Peptide Chain Termination, Translational; D010454:Peptide Termination Factors / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012343:RNA, Transfer / Q000494:pharmacology; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity; D016914:Ribonuclease H / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D020798:Two-Hybrid System Techniques; D014771:Virion / Q000378:metabolism; D014779:Virus Replication
C489003:Etf1 protein, mouse; D015685:Fusion Proteins, gag-pol; D010454:Peptide Termination Factors; D012343:RNA, Transfer; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase; D016914:Ribonuclease H