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. 1991 Aug 15;88(16):6991–6995. doi: 10.1073/pnas.88.16.6991

Evidence that a downstream pseudoknot is required for translational read-through of the Moloney murine leukemia virus gag stop codon.

N M Wills 1, R F Gesteland 1, J F Atkins 1
PMCID: PMC52219  PMID: 1871115

Abstract

Approximately 5% of the ribosomes translating the gag gene of murine leukemia viruses read through the UAG terminator and translate the in-frame pol gene to produce the gag-pol fusion polyprotein, the sole source of the pol gene products. We show that a pseudoknot located eight nucleotides 3' of the UAG codon in the Moloney murine leukemia virus is required for read-through. This requirement is markedly different from that known to be involved in other cases of read-through but surprisingly similar to some stimulatory sequences known to promote ribosomal frameshifting.

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Selected References

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  1. Atkins J. F., Gesteland R. F. Resolution of the discrepancy between a gene translation--termination codon and the deduced sequence for release of the encoded polypeptide. Eur J Biochem. 1983 Dec 15;137(3):509–516. doi: 10.1111/j.1432-1033.1983.tb07855.x. [DOI] [PubMed] [Google Scholar]
  2. Atkins J. F., Weiss R. B., Gesteland R. F. Ribosome gymnastics--degree of difficulty 9.5, style 10.0. Cell. 1990 Aug 10;62(3):413–423. doi: 10.1016/0092-8674(90)90007-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brierley I., Digard P., Inglis S. C. Characterization of an efficient coronavirus ribosomal frameshifting signal: requirement for an RNA pseudoknot. Cell. 1989 May 19;57(4):537–547. doi: 10.1016/0092-8674(89)90124-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown C. M., Stockwell P. A., Trotman C. N., Tate W. P. Sequence analysis suggests that tetra-nucleotides signal the termination of protein synthesis in eukaryotes. Nucleic Acids Res. 1990 Nov 11;18(21):6339–6345. doi: 10.1093/nar/18.21.6339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buckingham R. H., Sörensen P., Pagel F. T., Hijazi K. A., Mims B. H., Brechemier-Baey D., Murgola E. J. Third position base changes in codons 5' and 3' adjacent UGA codons affect UGA suppression in vivo. Biochim Biophys Acta. 1990 Aug 27;1050(1-3):259–262. doi: 10.1016/0167-4781(90)90177-4. [DOI] [PubMed] [Google Scholar]
  6. Capone J. P., Sedivy J. M., Sharp P. A., RajBhandary U. L. Introduction of UAG, UAA, and UGA nonsense mutations at a specific site in the Escherichia coli chloramphenicol acetyltransferase gene: use in measurement of amber, ochre, and opal suppression in mammalian cells. Mol Cell Biol. 1986 Sep;6(9):3059–3067. doi: 10.1128/mcb.6.9.3059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen E. Y., Seeburg P. H. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. doi: 10.1089/dna.1985.4.165. [DOI] [PubMed] [Google Scholar]
  8. Dinman J. D., Icho T., Wickner R. B. A -1 ribosomal frameshift in a double-stranded RNA virus of yeast forms a gag-pol fusion protein. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):174–178. doi: 10.1073/pnas.88.1.174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Felsenstein K. M., Goff S. P. Expression of the gag-pol fusion protein of Moloney murine leukemia virus without gag protein does not induce virion formation or proteolytic processing. J Virol. 1988 Jun;62(6):2179–2182. doi: 10.1128/jvi.62.6.2179-2182.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feng Y. X., Copeland T. D., Oroszlan S., Rein A., Levin J. G. Identification of amino acids inserted during suppression of UAA and UGA termination codons at the gag-pol junction of Moloney murine leukemia virus. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8860–8863. doi: 10.1073/pnas.87.22.8860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Feng Y. X., Levin J. G., Hatfield D. L., Schaefer T. S., Gorelick R. J., Rein A. Suppression of UAA and UGA termination codons in mutant murine leukemia viruses. J Virol. 1989 Jun;63(6):2870–2873. doi: 10.1128/jvi.63.6.2870-2873.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Herr W. Nucleotide sequence of AKV murine leukemia virus. J Virol. 1984 Feb;49(2):471–478. doi: 10.1128/jvi.49.2.471-478.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jacks T., Townsley K., Varmus H. E., Majors J. Two efficient ribosomal frameshifting events are required for synthesis of mouse mammary tumor virus gag-related polyproteins. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4298–4302. doi: 10.1073/pnas.84.12.4298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jones D. S., Nemoto F., Kuchino Y., Masuda M., Yoshikura H., Nishimura S. The effect of specific mutations at and around the gag-pol gene junction of Moloney murine leukaemia virus. Nucleic Acids Res. 1989 Aug 11;17(15):5933–5945. doi: 10.1093/nar/17.15.5933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kerr I. M., Olshevsky U., Lodish H. F., Baltimore D. Translation of murine leukemia virus RNA in cell-free systems from animal cells. J Virol. 1976 May;18(2):627–635. doi: 10.1128/jvi.18.2.627-635.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kuchino Y., Beier H., Akita N., Nishimura S. Natural UAG suppressor glutamine tRNA is elevated in mouse cells infected with Moloney murine leukemia virus. Proc Natl Acad Sci U S A. 1987 May;84(9):2668–2672. doi: 10.1073/pnas.84.9.2668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Murgola E. J., Hijazi K. A., Göringer H. U., Dahlberg A. E. Mutant 16S ribosomal RNA: a codon-specific translational suppressor. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4162–4165. doi: 10.1073/pnas.85.12.4162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murphy E. C., Jr, Arlinghaus R. B. Cell-free synthesis of Rauscher murine leukemia virus "gag" and "gag-pol" precursor polyproteins from virion 35 S RNA in a mRNA-dependent translation system derived from mouse tissue culture cells. Virology. 1978 May 15;86(2):329–343. doi: 10.1016/0042-6822(78)90074-0. [DOI] [PubMed] [Google Scholar]
  19. Murphy E. C., Jr, Wills N., Arlinghaus R. B. Suppression of murine retrovirus polypeptide termination: effect of amber suppressor tRNA on the cell-free translation of Rauscher murine leukemia virus, Moloney murine leukemia virus, and Moloney murine sarcoma virus 124 RNA. J Virol. 1980 May;34(2):464–473. doi: 10.1128/jvi.34.2.464-473.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Panganiban A. T. Retroviral gag gene amber codon suppression is caused by an intrinsic cis-acting component of the viral mRNA. J Virol. 1988 Oct;62(10):3574–3580. doi: 10.1128/jvi.62.10.3574-3580.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Philipson L., Andersson P., Olshevsky U., Weinberg R., Baltimore D., Gesteland R. Translation of MuLV and MSV RNAs in nuclease-treated reticulocyte extracts: enhancement of the gag-pol polypeptide with yeast suppressor tRNA. Cell. 1978 Jan;13(1):189–199. doi: 10.1016/0092-8674(78)90149-6. [DOI] [PubMed] [Google Scholar]
  22. Pleij C. W., Rietveld K., Bosch L. A new principle of RNA folding based on pseudoknotting. Nucleic Acids Res. 1985 Mar 11;13(5):1717–1731. doi: 10.1093/nar/13.5.1717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shinnick T. M., Lerner R. A., Sutcliffe J. G. Nucleotide sequence of Moloney murine leukaemia virus. Nature. 1981 Oct 15;293(5833):543–548. doi: 10.1038/293543a0. [DOI] [PubMed] [Google Scholar]
  24. Skuzeski J. M., Nichols L. M., Gesteland R. F., Atkins J. F. The signal for a leaky UAG stop codon in several plant viruses includes the two downstream codons. J Mol Biol. 1991 Mar 20;218(2):365–373. doi: 10.1016/0022-2836(91)90718-l. [DOI] [PubMed] [Google Scholar]
  25. Tang C. K., Draper D. E. Unusual mRNA pseudoknot structure is recognized by a protein translational repressor. Cell. 1989 May 19;57(4):531–536. doi: 10.1016/0092-8674(89)90123-2. [DOI] [PubMed] [Google Scholar]
  26. Weiss R. B., Dunn D. M., Atkins J. F., Gesteland R. F. Ribosomal frameshifting from -2 to +50 nucleotides. Prog Nucleic Acid Res Mol Biol. 1990;39:159–183. doi: 10.1016/s0079-6603(08)60626-1. [DOI] [PubMed] [Google Scholar]
  27. Weiss R. B., Huang W. M., Dunn D. M. A nascent peptide is required for ribosomal bypass of the coding gap in bacteriophage T4 gene 60. Cell. 1990 Jul 13;62(1):117–126. doi: 10.1016/0092-8674(90)90245-A. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wolin S. L., Walter P. Ribosome pausing and stacking during translation of a eukaryotic mRNA. EMBO J. 1988 Nov;7(11):3559–3569. doi: 10.1002/j.1460-2075.1988.tb03233.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wu H. N., Uhlenbeck O. C. Role of a bulged A residue in a specific RNA-protein interaction. Biochemistry. 1987 Dec 15;26(25):8221–8227. doi: 10.1021/bi00399a030. [DOI] [PubMed] [Google Scholar]
  30. Wyatt J. R., Puglisi J. D., Tinoco I., Jr RNA pseudoknots. Stability and loop size requirements. J Mol Biol. 1990 Jul 20;214(2):455–470. doi: 10.1016/0022-2836(90)90193-P. [DOI] [PubMed] [Google Scholar]
  31. Yoshinaka Y., Katoh I., Copeland T. D., Oroszlan S. Murine leukemia virus protease is encoded by the gag-pol gene and is synthesized through suppression of an amber termination codon. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1618–1622. doi: 10.1073/pnas.82.6.1618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yoshinaka Y., Katoh I., Copeland T. D., Oroszlan S. Translational readthrough of an amber termination codon during synthesis of feline leukemia virus protease. J Virol. 1985 Sep;55(3):870–873. doi: 10.1128/jvi.55.3.870-873.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zinoni F., Heider J., Böck A. Features of the formate dehydrogenase mRNA necessary for decoding of the UGA codon as selenocysteine. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4660–4664. doi: 10.1073/pnas.87.12.4660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. ten Dam E. B., Pleij C. W., Bosch L. RNA pseudoknots: translational frameshifting and readthrough on viral RNAs. Virus Genes. 1990 Jul;4(2):121–136. doi: 10.1007/BF00678404. [DOI] [PMC free article] [PubMed] [Google Scholar]

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