Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1989 Jul 25;17(14):5501–5507. doi: 10.1093/nar/17.14.5501

Translational reinitiation in the presence and absence of a Shine and Dalgarno sequence.

R A Spanjaard 1, J van Duin 1
PMCID: PMC318173  PMID: 2668889

Abstract

The process of translational reinitiation in Escherichia coli was studied in a two cistron system where expression of the downstream reporter gene was dependent on translation of an upstream reading frame. The dependence was almost absolute. Upstream translation increased expression of the downstream gene by two to three orders of magnitude. This large difference allowed us to quantitate restarts in a meaningful manner. In the absence of a Shine and Dalgarno (SD) region reinitiation occurred but its efficiency was about 10% of that found in the SD carrying counterpart. We discuss three ways by which translational coupling between neighboring cistrons can be enforced.

Full text

PDF

Images in this article

5504Image
on p.5504

Selected References

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

  1. Aksoy S., Squires C. L., Squires C. Translational coupling of the trpB and trpA genes in the Escherichia coli tryptophan operon. J Bacteriol. 1984 Feb;157(2):363–367. doi: 10.1128/jb.157.2.363-367.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berkhout B., Schmidt B. F., van Strien A., van Boom J., van Westrenen J., van Duin J. Lysis gene of bacteriophage MS2 is activated by translation termination at the overlapping coat gene. J Mol Biol. 1987 Jun 5;195(3):517–524. doi: 10.1016/0022-2836(87)90180-x. [DOI] [PubMed] [Google Scholar]
  3. Berkhout B., van der Laken C. J., van Knippenberg P. H. Formylmethionyl-tRNA binding to 30 S ribosomes programmed with homopolynucleotides and the effect of translational initiation factor 3. Biochim Biophys Acta. 1986 Mar 26;866(2-3):144–153. doi: 10.1016/0167-4781(86)90111-9. [DOI] [PubMed] [Google Scholar]
  4. Blumer K. J., Ivey M. R., Steege D. A. Translational control of phage f1 gene expression by differential activities of the gene V, VII, IX and VIII initiation sites. J Mol Biol. 1987 Oct 5;197(3):439–451. doi: 10.1016/0022-2836(87)90557-2. [DOI] [PubMed] [Google Scholar]
  5. Buell G., Schulz M. F., Selzer G., Chollet A., Movva N. R., Semon D., Escanez S., Kawashima E. Optimizing the expression in E. coli of a synthetic gene encoding somatomedin-C (IGF-I). Nucleic Acids Res. 1985 Mar 25;13(6):1923–1938. doi: 10.1093/nar/13.6.1923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cone K. C., Steege D. A. Messenger RNA conformation and ribosome selection of translational reinitiation sites in the lac repressor mRNA. J Mol Biol. 1985 Dec 20;186(4):725–732. doi: 10.1016/0022-2836(85)90392-4. [DOI] [PubMed] [Google Scholar]
  7. Das A., Yanofsky C. A ribosome binding site sequence is necessary for efficient expression of the distal gene of a translationally-coupled gene pair. Nucleic Acids Res. 1984 Jun 11;12(11):4757–4768. doi: 10.1093/nar/12.11.4757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dijkema R., Pouwels P., de Reus A., Schellekens H. Structure and expression in Escherichia coli of a cloned rat interferon-alpha gene. Nucleic Acids Res. 1984 Jan 25;12(2):1227–1242. doi: 10.1093/nar/12.2.1227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fiers W., Contreras R., Duerinck F., Haegeman G., Iserentant D., Merregaert J., Min Jou W., Molemans F., Raeymaekers A., Van den Berghe A. Complete nucleotide sequence of bacteriophage MS2 RNA: primary and secondary structure of the replicase gene. Nature. 1976 Apr 8;260(5551):500–507. doi: 10.1038/260500a0. [DOI] [PubMed] [Google Scholar]
  10. Hall M. N., Gabay J., Débarbouillé M., Schwartz M. A role for mRNA secondary structure in the control of translation initiation. Nature. 1982 Feb 18;295(5850):616–618. doi: 10.1038/295616a0. [DOI] [PubMed] [Google Scholar]
  11. Harms E., Higgins E., Chen J. W., Umbarger H. E. Translational coupling between the ilvD and ilvA genes of Escherichia coli. J Bacteriol. 1988 Oct;170(10):4798–4807. doi: 10.1128/jb.170.10.4798-4807.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Horinouchi S., Weisblum B. Posttranscriptional modification of mRNA conformation: mechanism that regulates erythromycin-induced resistance. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7079–7083. doi: 10.1073/pnas.77.12.7079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Looman A. C., Bodlaender J., de Gruyter M., Vogelaar A., van Knippenberg P. H. Secondary structure as primary determinant of the efficiency of ribosomal binding sites in Escherichia coli. Nucleic Acids Res. 1986 Jul 11;14(13):5481–5497. doi: 10.1093/nar/14.13.5481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Min Jou W., Haegeman G., Ysebaert M., Fiers W. Nucleotide sequence of the gene coding for the bacteriophage MS2 coat protein. Nature. 1972 May 12;237(5350):82–88. doi: 10.1038/237082a0. [DOI] [PubMed] [Google Scholar]
  15. Munson L. M., Stormo G. D., Niece R. L., Reznikoff W. S. lacZ translation initiation mutations. J Mol Biol. 1984 Aug 25;177(4):663–683. doi: 10.1016/0022-2836(84)90043-3. [DOI] [PubMed] [Google Scholar]
  16. Murialdo H., Siminovitch L. The morphogenesis of bacteriophage lambda. IV. Identification of gene products and control of the expression of the morphogenetic information. Virology. 1972 Jun;48(3):785–823. doi: 10.1016/0042-6822(72)90162-6. [DOI] [PubMed] [Google Scholar]
  17. Napoli C., Gold L., Singer B. S. Translational reinitiation in the rIIB cistron of bacteriophage T4. J Mol Biol. 1981 Jul 5;149(3):433–449. doi: 10.1016/0022-2836(81)90480-0. [DOI] [PubMed] [Google Scholar]
  18. Petersen H. U., Joseph E., Ullmann A., Danchin A. Formylation of initiator tRNA methionine in procaryotic protein synthesis: in vivo polarity in lactose operon expression. J Bacteriol. 1978 Aug;135(2):453–459. doi: 10.1128/jb.135.2.453-459.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Remaut E., Stanssens P., Fiers W. Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. Gene. 1981 Oct;15(1):81–93. doi: 10.1016/0378-1119(81)90106-2. [DOI] [PubMed] [Google Scholar]
  20. Schmidt B. F., Berkhout B., Overbeek G. P., van Strien A., van Duin J. Determination of the RNA secondary structure that regulates lysis gene expression in bacteriophage MS2. J Mol Biol. 1987 Jun 5;195(3):505–516. doi: 10.1016/0022-2836(87)90179-3. [DOI] [PubMed] [Google Scholar]
  21. Schoner B. E., Belagaje R. M., Schoner R. G. Translation of a synthetic two-cistron mRNA in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8506–8510. doi: 10.1073/pnas.83.22.8506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schümperli D., McKenney K., Sobieski D. A., Rosenberg M. Translational coupling at an intercistronic boundary of the Escherichia coli galactose operon. Cell. 1982 Oct;30(3):865–871. doi: 10.1016/0092-8674(82)90291-4. [DOI] [PubMed] [Google Scholar]
  23. Sor F., Bolotin-Fukuhara M., Nomura M. Mutational alterations of translational coupling in the L11 ribosomal protein operon of Escherichia coli. J Bacteriol. 1987 Aug;169(8):3495–3507. doi: 10.1128/jb.169.8.3495-3507.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Spanjaard R. A., van Dijk M. C., Turion A. J., van Duin J. Expression of the rat interferon-alpha 1 gene in Escherichia coli controlled by the secondary structure of the translation-initiation region. Gene. 1989 Aug 15;80(2):345–351. doi: 10.1016/0378-1119(89)90298-9. [DOI] [PubMed] [Google Scholar]
  25. Spanjaard R. A., van Duin J. Translation of the sequence AGG-AGG yields 50% ribosomal frameshift. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7967–7971. doi: 10.1073/pnas.85.21.7967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tessier L. H., Sondermeyer P., Faure T., Dreyer D., Benavente A., Villeval D., Courtney M., Lecocq J. P. The influence of mRNA primary and secondary structure on human IFN-gamma gene expression in E. coli. Nucleic Acids Res. 1984 Oct 25;12(20):7663–7675. doi: 10.1093/nar/12.20.7663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yates J. L., Nomura M. Feedback regulation of ribosomal protein synthesis in E. coli: localization of the mRNA target sites for repressor action of ribosomal protein L1. Cell. 1981 Apr;24(1):243–249. doi: 10.1016/0092-8674(81)90520-1. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES