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. 1995 Aug 1;14(15):3820–3827. doi: 10.1002/j.1460-2075.1995.tb00051.x

The Saccharomyces cerevisiae translation initiation factor Tif3 and its mammalian homologue, eIF-4B, have RNA annealing activity.

M Altmann 1, B Wittmer 1, N Méthot 1, N Sonenberg 1, H Trachsel 1
PMCID: PMC394456  PMID: 7543843

Abstract

The Saccharomyces cerevisiae TIF3 gene encodes the yeast homologue of mammalian translation initiation factor eIF-4B. We have added six histidine residues to the C-terminus of Tif3 protein (Tif3-His6p) and purified the tagged protein by affinity chromatography. Tif3-His6p stimulates translation and mRNA binding to ribosomes in a Tif3-dependent in vitro system. Furthermore, it binds to single-stranded RNA and catalyses the annealing of partially complementary RNA strands in vitro. In parallel experiments, RNA annealing activity could also be demonstrated for mammalian eIF-4B. A role for Tif3/eIF-4B and RNA annealing activity in the scanning process is proposed.

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Selected References

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

  1. Altmann M., Blum S., Wilson T. M., Trachsel H. 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. 1990 Jul 2;91(1):127–129. doi: 10.1016/0378-1119(90)90173-o. [DOI] [PubMed] [Google Scholar]
  2. Altmann M., Edery I., Sonenberg N., Trachsel H. Purification and characterization of protein synthesis initiation factor eIF-4E from the yeast Saccharomyces cerevisiae. Biochemistry. 1985 Oct 22;24(22):6085–6089. doi: 10.1021/bi00343a009. [DOI] [PubMed] [Google Scholar]
  3. Altmann M., Edery I., Trachsel H., Sonenberg N. Site-directed mutagenesis of the tryptophan residues in yeast eukaryotic initiation factor 4E. Effects on cap binding activity. J Biol Chem. 1988 Nov 25;263(33):17229–17232. [PubMed] [Google Scholar]
  4. Altmann M., Müller P. P., Wittmer B., Ruchti F., Lanker S., Trachsel H. A Saccharomyces cerevisiae homologue of mammalian translation initiation factor 4B contributes to RNA helicase activity. EMBO J. 1993 Oct;12(10):3997–4003. doi: 10.1002/j.1460-2075.1993.tb06077.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Altmann M., Sonenberg N., Trachsel H. Translation in Saccharomyces cerevisiae: initiation factor 4E-dependent cell-free system. Mol Cell Biol. 1989 Oct;9(10):4467–4472. doi: 10.1128/mcb.9.10.4467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Altmann M., Trachsel H. Regulation of translation initiation and modulation of cellular physiology. Trends Biochem Sci. 1993 Nov;18(11):429–432. doi: 10.1016/0968-0004(93)90143-b. [DOI] [PubMed] [Google Scholar]
  7. Anderson C. W., Baum P. R., Gesteland R. F. Processing of adenovirus 2-induced proteins. J Virol. 1973 Aug;12(2):241–252. doi: 10.1128/jvi.12.2.241-252.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Biamonti G., Riva S. New insights into the auxiliary domains of eukaryotic RNA binding proteins. FEBS Lett. 1994 Feb 28;340(1-2):1–8. doi: 10.1016/0014-5793(94)80162-2. [DOI] [PubMed] [Google Scholar]
  9. Blum S., Mueller M., Schmid S. R., Linder P., Trachsel H. Translation in Saccharomyces cerevisiae: initiation factor 4A-dependent cell-free system. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6043–6046. doi: 10.1073/pnas.86.16.6043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Blum S., Schmid S. R., Pause A., Buser P., Linder P., Sonenberg N., Trachsel H. ATP hydrolysis by initiation factor 4A is required for translation initiation in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7664–7668. doi: 10.1073/pnas.89.16.7664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Boeck R., Kolakofsky D. Positions +5 and +6 can be major determinants of the efficiency of non-AUG initiation codons for protein synthesis. EMBO J. 1994 Aug 1;13(15):3608–3617. doi: 10.1002/j.1460-2075.1994.tb06668.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chien C. T., Bartel P. L., Sternglanz R., Fields S. The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9578–9582. doi: 10.1073/pnas.88.21.9578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cobianchi F., Calvio C., Stoppini M., Buvoli M., Riva S. Phosphorylation of human hnRNP protein A1 abrogates in vitro strand annealing activity. Nucleic Acids Res. 1993 Feb 25;21(4):949–955. doi: 10.1093/nar/21.4.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Coppolecchia R., Buser P., Stotz A., Linder P. A new yeast translation initiation factor suppresses a mutation in the eIF-4A RNA helicase. EMBO J. 1993 Oct;12(10):4005–4011. doi: 10.1002/j.1460-2075.1993.tb06078.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Duncan R., Hershey J. W. Identification and quantitation of levels of protein synthesis initiation factors in crude HeLa cell lysates by two-dimensional polyacrylamide gel electrophoresis. J Biol Chem. 1983 Jun 10;258(11):7228–7235. [PubMed] [Google Scholar]
  16. Grünert S., Jackson R. J. The immediate downstream codon strongly influences the efficiency of utilization of eukaryotic translation initiation codons. EMBO J. 1994 Aug 1;13(15):3618–3630. doi: 10.1002/j.1460-2075.1994.tb06669.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hershey J. W. Translational control in mammalian cells. Annu Rev Biochem. 1991;60:717–755. doi: 10.1146/annurev.bi.60.070191.003441. [DOI] [PubMed] [Google Scholar]
  18. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kenan D. J., Query C. C., Keene J. D. RNA recognition: towards identifying determinants of specificity. Trends Biochem Sci. 1991 Jun;16(6):214–220. doi: 10.1016/0968-0004(91)90088-d. [DOI] [PubMed] [Google Scholar]
  20. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Lagúnez-Otero J. rRNA-mRNA complementarity: implications for translation initiation. Trends Biochem Sci. 1993 Nov;18(11):406–408. doi: 10.1016/0968-0004(93)90134-9. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Milburn S. C., Hershey J. W., Davies M. V., Kelleher K., Kaufman R. J. Cloning and expression of eukaryotic initiation factor 4B cDNA: sequence determination identifies a common RNA recognition motif. EMBO J. 1990 Sep;9(9):2783–2790. doi: 10.1002/j.1460-2075.1990.tb07466.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Méthot N., Pause A., Hershey J. W., Sonenberg N. The translation initiation factor eIF-4B contains an RNA-binding region that is distinct and independent from its ribonucleoprotein consensus sequence. Mol Cell Biol. 1994 Apr;14(4):2307–2316. doi: 10.1128/mcb.14.4.2307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Naranda T., Strong W. B., Menaya J., Fabbri B. J., Hershey J. W. Two structural domains of initiation factor eIF-4B are involved in binding to RNA. J Biol Chem. 1994 May 20;269(20):14465–14472. [PubMed] [Google Scholar]
  27. Oberosler P., Hloch P., Ramsperger U., Stahl H. p53-catalyzed annealing of complementary single-stranded nucleic acids. EMBO J. 1993 Jun;12(6):2389–2396. doi: 10.1002/j.1460-2075.1993.tb05893.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pause A., Sonenberg N. Mutational analysis of a DEAD box RNA helicase: the mammalian translation initiation factor eIF-4A. EMBO J. 1992 Jul;11(7):2643–2654. doi: 10.1002/j.1460-2075.1992.tb05330.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Portman D. S., Dreyfuss G. RNA annealing activities in HeLa nuclei. EMBO J. 1994 Jan 1;13(1):213–221. doi: 10.1002/j.1460-2075.1994.tb06251.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rhoads R. E. Regulation of eukaryotic protein synthesis by initiation factors. J Biol Chem. 1993 Feb 15;268(5):3017–3020. [PubMed] [Google Scholar]
  31. Rozen F., Edery I., Meerovitch K., Dever T. E., Merrick W. C., Sonenberg N. Bidirectional RNA helicase activity of eucaryotic translation initiation factors 4A and 4F. Mol Cell Biol. 1990 Mar;10(3):1134–1144. doi: 10.1128/mcb.10.3.1134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Sonenberg N. Translation factors as effectors of cell growth and tumorigenesis. Curr Opin Cell Biol. 1993 Dec;5(6):955–960. doi: 10.1016/0955-0674(93)90076-3. [DOI] [PubMed] [Google Scholar]
  34. 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]

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