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. 1995 Mar 11;23(5):861–868. doi: 10.1093/nar/23.5.861

The first and third uORFs in RSV leader RNA are efficiently translated: implications for translational regulation and viral RNA packaging.

O Donzé 1, P Damay 1, P F Spahr 1
PMCID: PMC306771  PMID: 7708504

Abstract

Rous sarcoma virus (RSV) RNA leader contains three short upstream open reading frames. We have shown recently that both uORFs 1 and 3 influence in vivo translation of the downstream gag gene and are involved in the virus RNA packaging process. In this report, we have studied the translational events occurring at the upstream AUGs in vivo. We show that (i) the first and third AUGs are efficient translational initiation sites; (ii) ribosomes reinitiate efficiently at AUG3; and (iii) deletions in the intercistronic distance between uORF1 and 3 (which is well conserved among avian strains) prevent ribosome initiation at AUG3, thus increasing translation efficiency at the downstream AUGgag. The roles of the uORFs in translation and packaging are discussed.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abastado J. P., Miller P. F., Jackson B. M., Hinnebusch A. G. Suppression of ribosomal reinitiation at upstream open reading frames in amino acid-starved cells forms the basis for GCN4 translational control. Mol Cell Biol. 1991 Jan;11(1):486–496. doi: 10.1128/mcb.11.1.486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aronoff R., Hajjar A. M., Linial M. L. Avian retroviral RNA encapsidation: reexamination of functional 5' RNA sequences and the role of nucleocapsid Cys-His motifs. J Virol. 1993 Jan;67(1):178–188. doi: 10.1128/jvi.67.1.178-188.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bizub D., Katz R. A., Skalka A. M. Nucleotide sequence of noncoding regions in Rous-associated virus-2: comparisons delineate conserved regions important in replication and oncogenesis. J Virol. 1984 Feb;49(2):557–565. doi: 10.1128/jvi.49.2.557-565.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cobrinik D., Aiyar A., Ge Z., Katzman M., Huang H., Leis J. Overlapping retrovirus U5 sequence elements are required for efficient integration and initiation of reverse transcription. J Virol. 1991 Jul;65(7):3864–3872. doi: 10.1128/jvi.65.7.3864-3872.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cobrinik D., Soskey L., Leis J. A retroviral RNA secondary structure required for efficient initiation of reverse transcription. J Virol. 1988 Oct;62(10):3622–3630. doi: 10.1128/jvi.62.10.3622-3630.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Damiani R. D., Jr, Wessler S. R. An upstream open reading frame represses expression of Lc, a member of the R/B family of maize transcriptional activators. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8244–8248. doi: 10.1073/pnas.90.17.8244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Darlix J. L., Spahr P. F., Bromley P. A., Jaton J. C. In vitro, the major ribosome binding site on Rous sarcoma virus RNA does not contain the nucleotide sequence coding for the N-terminal amino acids of the gag gene product. J Virol. 1979 Feb;29(2):597–611. doi: 10.1128/jvi.29.2.597-611.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Degnin C. R., Schleiss M. R., Cao J., Geballe A. P. Translational inhibition mediated by a short upstream open reading frame in the human cytomegalovirus gpUL4 (gp48) transcript. J Virol. 1993 Sep;67(9):5514–5521. doi: 10.1128/jvi.67.9.5514-5521.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Donzé O., Spahr P. F. Role of the open reading frames of Rous sarcoma virus leader RNA in translation and genome packaging. EMBO J. 1992 Oct;11(10):3747–3757. doi: 10.1002/j.1460-2075.1992.tb05460.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fütterer J., Hohn T. Role of an upstream open reading frame in the translation of polycistronic mRNAs in plant cells. Nucleic Acids Res. 1992 Aug 11;20(15):3851–3857. doi: 10.1093/nar/20.15.3851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fütterer J., Hohn T. Translation of a polycistronic mRNA in the presence of the cauliflower mosaic virus transactivator protein. EMBO J. 1991 Dec;10(12):3887–3896. doi: 10.1002/j.1460-2075.1991.tb04958.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Grant C. M., Hinnebusch A. G. Effect of sequence context at stop codons on efficiency of reinitiation in GCN4 translational control. Mol Cell Biol. 1994 Jan;14(1):606–618. doi: 10.1128/mcb.14.1.606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hackett P. B., Dalton M. W., Johnson D. P., Petersen R. B. Phylogenetic and physical analysis of the 5' leader RNA sequences of avian retroviruses. Nucleic Acids Res. 1991 Dec 25;19(24):6929–6934. doi: 10.1093/nar/19.24.6929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hackett P. B., Petersen R. B., Hensel C. H., Albericio F., Gunderson S. I., Palmenberg A. C., Barany G. Synthesis in vitro of a seven amino acid peptide encoded in the leader RNA of Rous sarcoma virus. J Mol Biol. 1986 Jul 5;190(1):45–57. doi: 10.1016/0022-2836(86)90074-4. [DOI] [PubMed] [Google Scholar]
  15. Hensel C. H., Petersen R. B., Hackett P. B. Effects of alterations in the leader sequence of Rous sarcoma virus RNA on initiation of translation. J Virol. 1989 Nov;63(11):4986–4990. doi: 10.1128/jvi.63.11.4986-4990.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hill J. R., Morris D. R. Cell-specific translational regulation of S-adenosylmethionine decarboxylase mRNA. Dependence on translation and coding capacity of the cis-acting upstream open reading frame. J Biol Chem. 1993 Jan 5;268(1):726–731. [PubMed] [Google Scholar]
  17. Hinnebusch A. G., Jackson B. M., Mueller P. P. Evidence for regulation of reinitiation in translational control of GCN4 mRNA. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7279–7283. doi: 10.1073/pnas.85.19.7279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hughes S., Mellstrom K., Kosik E., Tamanoi F., Brugge J. Mutation of a termination codon affects src initiation. Mol Cell Biol. 1984 Sep;4(9):1738–1746. doi: 10.1128/mcb.4.9.1738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Katz R. A., Terry R. W., Skalka A. M. A conserved cis-acting sequence in the 5' leader of avian sarcoma virus RNA is required for packaging. J Virol. 1986 Jul;59(1):163–167. doi: 10.1128/jvi.59.1.163-167.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kawai S., Koyama T. Characterization of a Rous sarcoma virus mutant defective in packaging its own genomic RNA: biological properties of mutant TK15 and mutant-induced transformants. J Virol. 1984 Jul;51(1):147–153. doi: 10.1128/jvi.51.1.147-153.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Knight J. B., Si Z. H., Stoltzfus C. M. A base-paired structure in the avian sarcoma virus 5' leader is required for efficient encapsidation of RNA. J Virol. 1994 Jul;68(7):4493–4502. doi: 10.1128/jvi.68.7.4493-4502.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kozak M. An analysis of vertebrate mRNA sequences: intimations of translational control. J Cell Biol. 1991 Nov;115(4):887–903. doi: 10.1083/jcb.115.4.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kozak M. Effects of intercistronic length on the efficiency of reinitiation by eucaryotic ribosomes. Mol Cell Biol. 1987 Oct;7(10):3438–3445. doi: 10.1128/mcb.7.10.3438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kozak M. Influences of mRNA secondary structure on initiation by eukaryotic ribosomes. Proc Natl Acad Sci U S A. 1986 May;83(9):2850–2854. doi: 10.1073/pnas.83.9.2850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Linial M. L., Miller A. D. Retroviral RNA packaging: sequence requirements and implications. Curr Top Microbiol Immunol. 1990;157:125–152. doi: 10.1007/978-3-642-75218-6_5. [DOI] [PubMed] [Google Scholar]
  28. Lo K. M., Jones S. S., Hackett N. R., Khorana H. G. Specific amino acid substitutions in bacterioopsin: Replacement of a restriction fragment in the structural gene by synthetic DNA fragments containing altered codons. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2285–2289. doi: 10.1073/pnas.81.8.2285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Merrick W. C. Mechanism and regulation of eukaryotic protein synthesis. Microbiol Rev. 1992 Jun;56(2):291–315. doi: 10.1128/mr.56.2.291-315.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Miller P. F., Hinnebusch A. G. Sequences that surround the stop codons of upstream open reading frames in GCN4 mRNA determine their distinct functions in translational control. Genes Dev. 1989 Aug;3(8):1217–1225. doi: 10.1101/gad.3.8.1217. [DOI] [PubMed] [Google Scholar]
  31. Moustakas A., Sonstegard T. S., Hackett P. B. Alterations of the three short open reading frames in the Rous sarcoma virus leader RNA modulate viral replication and gene expression. J Virol. 1993 Jul;67(7):4337–4349. doi: 10.1128/jvi.67.7.4337-4349.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Moustakas A., Sonstegard T. S., Hackett P. B. Effects of the open reading frames in the Rous sarcoma virus leader RNA on translation. J Virol. 1993 Jul;67(7):4350–4357. doi: 10.1128/jvi.67.7.4350-4357.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mueller P. P., Jackson B. M., Miller P. F., Hinnebusch A. G. The first and fourth upstream open reading frames in GCN4 mRNA have similar initiation efficiencies but respond differently in translational control to change in length and sequence. Mol Cell Biol. 1988 Dec;8(12):5439–5447. doi: 10.1128/mcb.8.12.5439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Méric C., Spahr P. F. Rous sarcoma virus nucleic acid-binding protein p12 is necessary for viral 70S RNA dimer formation and packaging. J Virol. 1986 Nov;60(2):450–459. doi: 10.1128/jvi.60.2.450-459.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Oertle S., Spahr P. F. Role of the gag polyprotein precursor in packaging and maturation of Rous sarcoma virus genomic RNA. J Virol. 1990 Dec;64(12):5757–5763. doi: 10.1128/jvi.64.12.5757-5763.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Peabody D. S., Berg P. Termination-reinitiation occurs in the translation of mammalian cell mRNAs. Mol Cell Biol. 1986 Jul;6(7):2695–2703. doi: 10.1128/mcb.6.7.2695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Pelletier J., Sonenberg N. Insertion mutagenesis to increase secondary structure within the 5' noncoding region of a eukaryotic mRNA reduces translational efficiency. Cell. 1985 Mar;40(3):515–526. doi: 10.1016/0092-8674(85)90200-4. [DOI] [PubMed] [Google Scholar]
  38. Pelletier J., Sonenberg N. Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA. Nature. 1988 Jul 28;334(6180):320–325. doi: 10.1038/334320a0. [DOI] [PubMed] [Google Scholar]
  39. Petersen R. B., Hackett P. B. Characterization of ribosome binding on Rous sarcoma virus RNA in vitro. J Virol. 1985 Dec;56(3):683–690. doi: 10.1128/jvi.56.3.683-690.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Petersen R. B., Hensel C. H., Hackett P. B. Identification of a ribosome-binding site for a leader peptide encoded by Rous sarcoma virus RNA. J Virol. 1984 Sep;51(3):722–729. doi: 10.1128/jvi.51.3.722-729.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Petersen R. B., Moustakas A., Hackett P. B. A mutation in the short 5'-proximal open reading frame on Rous sarcoma virus RNA alters virus production. J Virol. 1989 Nov;63(11):4787–4796. doi: 10.1128/jvi.63.11.4787-4796.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Werner M., Feller A., Messenguy F., Piérard A. The leader peptide of yeast gene CPA1 is essential for the translational repression of its expression. Cell. 1987 Jun 19;49(6):805–813. doi: 10.1016/0092-8674(87)90618-0. [DOI] [PubMed] [Google Scholar]
  43. de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]

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