Summary
Synthetic oligonucleotides encoding the 5′-non-translated (leader) sequence of the coat protein mRNA of alfalfa mosaic virus RNA 4 or the Ω′ leader sequence of tobacco mosaic virus RNA were used to replace the natural leader region of the yeast phosphoglycerate kinase (PGK1) mRNAs and the translational efficiency of the chimeric mRNA was determined in yeast cells. In neither case did we observed a significant increase compared to the translational efficiency shown by the wild-type PGK mRNA, in contrast to the known stimulatory effect of these leader sequences on translation in mammalian, plant and bacterial in-vivo and/or in-vitro systems. The same result was obtained when the translational efficiencies in yeast cells of Escherichia coli β-galactosidase mRNAs carrying the PGK or either of the two viral leader sequences were compared.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Altmann M, Sonenberg N, Trachsel H (1989) Translation in Saccharomyces cerevisiae: initiation factor 4E-dependent cell-free system. Mol Cell Biol 9:4467–4472
Altmann M, Blum S, Wilson TMA, Trachsel H (1990) The 5′-leader sequence of tobacco mosaic virus RNA mediates initiation-factor-4E-independent, but still initiation-factor-4A-dependent translation in yeast extracts. Gene 91:127–129
Barbu V, Dautry F (1989) Northern blot normalization with a 28S rRNA oligonucleotide probe. Nucleic Acids Res 17:7115
Casadaban MJ, Martinez-Arias A, Shapira SK, Chou J (1983) β-Galatosidase gene fusions for analyzing gene expression in Escherichia coli and yeast. Methods Enzymol 100:293–308
Gallie DR, Kado CI (1989) A translational enhancer derived from tobacco mosaic virus is functionally equivalent to a Shine-Dalgarno sequence. Proc Natl Acad Sci USA 86:129–132
Gallie DR, Sleat DE, Watts JW, Turner PC, Wilson TMA (1987a) The 5′-leader sequence of tobacco mosaic virus RNA enhances the expression of foreign gene transcripts in vitro and in vivo. Nucleic Acids Res 15:3257–3273
Gallie DR, Sleat DE, Watts JW, Turner PC, Wilson TMA (1987b) A comparison of eukaryotic viral 5′-leader sequences as enhancers of mRNA expression in vivo. Nucleic Acids Res 15:8693–8711
Gallie DR, Walbot V, Hershey JWB (1988) The ribosomal fraction mediates the translational enhancement associated with the 5′-leader of tobacco mosaic virus. Nucleic Acids Res 16:8675–8694
Gehrke L, Jobling SA (1990) Untranslated leader sequences and enhanced messenger RNA translational efficiency. In: McCarthy JEG, Tuite MF (eds) Post-transcriptional control of gene expression. Springer, Berlin Heidelberg New York, p 389–398
Gietz RD, Sugino A (1988) New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base-pair restriction sites. Gene 74:527–534
Heuvel JJ van den, Bergkamp RJM, Planta RJ, Raué HA (1989) Effect of deletions in the 5′-noncoding region on the translational efficiency of phosphoglycerate kinase mRNA in yeast. Gene 79:83–95
Heuvel JJ van den, Planta RJ, Raué HA (1990) Effect of leader primary structure on the translational efficiency of phosphoglycerate kinase mRNA in yeast. Yeast 6:473–482
Hoekema A, Kastelein RA, Vasser M, Boer HA de (1987) Codon replacement in the PGK1 gene of Saccharomyces cerevisiae: experimental approach to study the role of biased codon usage in gene expression. Mol Cell Biol 7:2914–2924
Jobling SA, Gehrke L (1987) Enhanced translation of chimaeric messenger RNAs containing a plant viral untranslated leader sequence. Nature 325:622–625
Jobling SA, Cuthbert CM, Rogers SG, Fraley RT, Gehrke L (1988) In vitro transcription and translational efficiency of chimeric SP6 messenger RNAs devoid of 5′ vector nucleotides. Nucleic Acids Res 16:4483–4498
Jones E (1976) Proteinase mutants of Saccharomyces cerevisiae. Genetics 85:23–33
Kaiser CA, Botstein D (1986) Secretion-defective mutations in the signal sequence for Saccharomyces cerevisiae invertase. Mol Cell Biol 6:2382–2391
Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227:680–685
Mellor J, Dobson MJ, Roberts NA, Kingsman AJ, Kingsman S (1985) Factors affecting heterologous gene expression in Saccharomyces cerevisiae. Gene 33:215–226
Miller JH (1972) Experiments in molecular genetics, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
Raué HA, Heuvel JJ van den, Planta RJ (1990) Yeast mRNA structure and translational efficiency. In: McCarthy JEG, Tuite MF (eds) Post-transcriptional control of gene expression. Springer, Berlin Heidelberg New York, p 237–247
Sleat DE, Gallie DR, Jefferson RA, Bevan MW, Turner PC, Wilson TMA (1987) Characterization of the 5′-leader sequence of tobacco mosaic virus RNA as a general enhancer of translation in vitro. Gene 217:217–225
Sleat DE, Hull R, Turner PC, Wilson TMA (1988) Studies on the mechanism of translational enhancement by the 5′-leader sequence of tobacco mosaic virus RNA. Eur J Biochem 175:75–86
Wilson TMA, Saunders K, Dowson-Day M, Sleat DE, Trachsel H, Mundry KW (1990) Effects of the 5′-leader sequence of tobacco mosaic virus RNA, or derivatives thereof on foreign mRNA and native viral gene expression. In: McCarthy JEG, Tuite MF (eds) Post-transcriptional control of gene expression. Springer, Berlin Heidelberg New York, p 261–275
Author information
Authors and Affiliations
Additional information
Offprint requests to: H. A. Raué
Rights and permissions
About this article
Cite this article
van der Heuvel, J.J., Raué, H.A. Translational efficiency of mRNA in yeast cells is not increased by plant viral leader sequences. Appl Microbiol Biotechnol 37, 84–87 (1992). https://doi.org/10.1007/BF00174208
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00174208