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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1989 Jan;86(1):129–132. doi: 10.1073/pnas.86.1.129

A translational enhancer derived from tobacco mosaic virus is functionally equivalent to a Shine-Dalgarno sequence.

D R Gallie 1, C I Kado 1
PMCID: PMC286417  PMID: 2643095

Abstract

When present at the 5' end of mRNAs, the untranslated leader sequence (omega) of tobacco mosaic virus RNA significantly enhances translation in eukaryotes and prokaryotes. We have tested a deletion derivative of the omega sequence, omega delta 3, for its enhancing ability on gene constructs in which the ribosomal binding site was either present or deleted, in several Gram-negative bacterial species including Escherichia coli, Agrobacterium tumefaciens, Xanthomonas campestris pv. vitians, Erwinia amylovora, and Salmonella typhimurium. In vivo production of chloramphenicol acetyltransferase from a gene construct lacking its native ribosomal binding site was enhanced 40- to 120-fold by the presence of omega delta 3. Similar levels of enhancement (30- to 240-fold) were observed when the gene encoding beta-glucuronidase was tested. With a chloramphenicol acetyltransferase construct containing a ribosomal binding site, enhancement was markedly less, between 1- and 3.8-fold. Omega delta 3 appeared to enhance translation independent of its position upstream of the AUG codon used for initiation.

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Selected References

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

  1. Belasco J. G., Nilsson G., von Gabain A., Cohen S. N. The stability of E. coli gene transcripts is dependent on determinants localized to specific mRNA segments. Cell. 1986 Jul 18;46(2):245–251. doi: 10.1016/0092-8674(86)90741-5. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Close T. J., Rodriguez R. L. Construction and characterization of the chloramphenicol-resistance gene cartridge: a new approach to the transcriptional mapping of extrachromosomal elements. Gene. 1982 Dec;20(2):305–316. doi: 10.1016/0378-1119(82)90048-8. [DOI] [PubMed] [Google Scholar]
  4. Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. A comparison of eukaryotic viral 5'-leader sequences as enhancers of mRNA expression in vivo. Nucleic Acids Res. 1987 Nov 11;15(21):8693–8711. doi: 10.1093/nar/15.21.8693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. In vivo uncoating and efficient expression of foreign mRNAs packaged in TMV-like particles. Science. 1987 May 29;236(4805):1122–1124. doi: 10.1126/science.3472350. [DOI] [PubMed] [Google Scholar]
  6. Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. Mutational analysis of the tobacco mosaic virus 5'-leader for altered ability to enhance translation. Nucleic Acids Res. 1988 Feb 11;16(3):883–893. doi: 10.1093/nar/16.3.883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. The 5'-leader sequence of tobacco mosaic virus RNA enhances the expression of foreign gene transcripts in vitro and in vivo. Nucleic Acids Res. 1987 Apr 24;15(8):3257–3273. doi: 10.1093/nar/15.8.3257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gallie D. R., Walbot V., Hershey J. W. The ribosomal fraction mediates the translational enhancement associated with the 5'-leader of tobacco mosaic virus. Nucleic Acids Res. 1988 Sep 12;16(17):8675–8694. doi: 10.1093/nar/16.17.8675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ganoza M. C., Kofoid E. C., Marlière P., Louis B. G. Potential secondary structure at translation-initiation sites. Nucleic Acids Res. 1987 Jan 12;15(1):345–360. doi: 10.1093/nar/15.1.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gheysen D., Iserentant D., Derom C., Fiers W. Systematic alteration of the nucleotide sequence preceding the translation initiation codon and the effects on bacterial expression of the cloned SV40 small-t antigen gene. Gene. 1982 Jan;17(1):55–63. doi: 10.1016/0378-1119(82)90100-7. [DOI] [PubMed] [Google Scholar]
  11. Gold L., Pribnow D., Schneider T., Shinedling S., Singer B. S., Stormo G. Translational initiation in prokaryotes. Annu Rev Microbiol. 1981;35:365–403. doi: 10.1146/annurev.mi.35.100181.002053. [DOI] [PubMed] [Google Scholar]
  12. Gold L., Stormo G., Saunders R. Escherichia coli translational initiation factor IF3: a unique case of translational regulation. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7061–7065. doi: 10.1073/pnas.81.22.7061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jay G., Khoury G., Seth A. K., Jay E. Construction of a general vector for efficient expression of mammalian proteins in bacteria: use of a synthetic ribosome binding site. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5543–5548. doi: 10.1073/pnas.78.9.5543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jefferson R. A., Burgess S. M., Hirsh D. beta-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8447–8451. doi: 10.1073/pnas.83.22.8447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morrison D. A. Transformation and preservation of competent bacterial cells by freezing. Methods Enzymol. 1979;68:326–331. doi: 10.1016/0076-6879(79)68023-0. [DOI] [PubMed] [Google Scholar]
  17. Pirrotta V. Operators and promoters in the OR region of phage 434. Nucleic Acids Res. 1979 Apr;6(4):1495–1508. doi: 10.1093/nar/6.4.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ptashne M., Backman K., Humayun M. Z., Jeffrey A., Maurer R., Meyer B., Sauer R. T. Autoregulation and function of a repressor in bacteriophage lambda. Science. 1976 Oct 8;194(4261):156–161. doi: 10.1126/science.959843. [DOI] [PubMed] [Google Scholar]
  19. Roberts T. M., Kacich R., Ptashne M. A general method for maximizing the expression of a cloned gene. Proc Natl Acad Sci U S A. 1979 Feb;76(2):760–764. doi: 10.1073/pnas.76.2.760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rogowsky P. M., Close T. J., Chimera J. A., Shaw J. J., Kado C. I. Regulation of the vir genes of Agrobacterium tumefaciens plasmid pTiC58. J Bacteriol. 1987 Nov;169(11):5101–5112. doi: 10.1128/jb.169.11.5101-5112.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schneider E., Blundell M., Kennell D. Translation and mRNA decay. Mol Gen Genet. 1978 Apr 6;160(2):121–129. doi: 10.1007/BF00267473. [DOI] [PubMed] [Google Scholar]
  22. Shepard H. M., Yelverton E., Goeddel D. V. Increased synthesis in E. coli of fibroblast and leukocyte interferons through alterations in ribosome binding sites. DNA. 1982;1(2):125–131. doi: 10.1089/dna.1.1982.1.125. [DOI] [PubMed] [Google Scholar]
  23. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sleat D. E., Gallie D. R., Jefferson R. A., Bevan M. W., Turner P. C., Wilson T. M. Characterisation of the 5'-leader sequence of tobacco mosaic virus RNA as a general enhancer of translation in vitro. Gene. 1987;60(2-3):217–225. doi: 10.1016/0378-1119(87)90230-7. [DOI] [PubMed] [Google Scholar]
  25. Sleat D. E., Hull R., Turner P. C., Wilson T. M. Studies on the mechanism of translational enhancement by the 5'-leader sequence of tobacco mosaic virus RNA. Eur J Biochem. 1988 Jul 15;175(1):75–86. doi: 10.1111/j.1432-1033.1988.tb14168.x. [DOI] [PubMed] [Google Scholar]
  26. Smiley B. L., Lupski J. R., Svec P. S., McMacken R., Godson G. N. Sequences of the Escherichia coli dnaG primase gene and regulation of its expression. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4550–4554. doi: 10.1073/pnas.79.15.4550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Steitz J. A., Jakes K. How ribosomes select initiator regions in mRNA: base pair formation between the 3' terminus of 16S rRNA and the mRNA during initiation of protein synthesis in Escherichia coli. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4734–4738. doi: 10.1073/pnas.72.12.4734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stormo G. D., Schneider T. D., Gold L. M. Characterization of translational initiation sites in E. coli. Nucleic Acids Res. 1982 May 11;10(9):2971–2996. doi: 10.1093/nar/10.9.2971. [DOI] [PMC free article] [PubMed] [Google Scholar]

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