Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1993 Oct;12(10):4005–4011. doi: 10.1002/j.1460-2075.1993.tb06078.x

A new yeast translation initiation factor suppresses a mutation in the eIF-4A RNA helicase.

R Coppolecchia 1, P Buser 1, A Stotz 1, P Linder 1
PMCID: PMC413683  PMID: 8404866

Abstract

We have isolated a gene, STM1, which encodes a new translation initiation factor from Saccharomyces cerevisiae. The gene acts, if present on a multicopy plasmid, as a suppressor of a temperature-sensitive mutation in eIF-4A. The single copy STM1 gene is not essential, but disruption causes a slow growth phenotype. Analysis of polysomes from a strain carrying a disrupted stm1 allele shows a clear defect in translation initiation as shown by a strong reduction in polysomes and an increase in the monosomes. Sequence analysis revealed interesting features of the putative Stm1 protein. Comparison of the entire protein sequence with databanks showed some similarity with the human eIF-4B protein. The Stm1 protein has potential RNP1 and RNP2 motifs characteristic for RNA-binding proteins. The protein also contains six highly conserved direct repeats of 21-26 amino acids and one partial repeat.

Full text

PDF
4005

Images in this article

4007Image
on p.4007
4009Image
on p.4009

Selected References

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

  1. Abramson R. D., Dever T. E., Lawson T. G., Ray B. K., Thach R. E., Merrick W. C. The ATP-dependent interaction of eukaryotic initiation factors with mRNA. J Biol Chem. 1987 Mar 15;262(8):3826–3832. [PubMed] [Google Scholar]
  2. 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]
  3. Baim S. B., Pietras D. F., Eustice D. C., Sherman F. A mutation allowing an mRNA secondary structure diminishes translation of Saccharomyces cerevisiae iso-1-cytochrome c. Mol Cell Biol. 1985 Aug;5(8):1839–1846. doi: 10.1128/mcb.5.8.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Banroques J., Delahodde A., Jacq C. A mitochondrial RNA maturase gene transferred to the yeast nucleus can control mitochondrial mRNA splicing. Cell. 1986 Sep 12;46(6):837–844. doi: 10.1016/0092-8674(86)90065-6. [DOI] [PubMed] [Google Scholar]
  5. Bennetzen J. L., Hall B. D. Codon selection in yeast. J Biol Chem. 1982 Mar 25;257(6):3026–3031. [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Bonneaud N., Ozier-Kalogeropoulos O., Li G. Y., Labouesse M., Minvielle-Sebastia L., Lacroute F. A family of low and high copy replicative, integrative and single-stranded S. cerevisiae/E. coli shuttle vectors. Yeast. 1991 Aug-Sep;7(6):609–615. doi: 10.1002/yea.320070609. [DOI] [PubMed] [Google Scholar]
  9. Browning K. S., Fletcher L., Lax S. R., Ravel J. M. Evidence that the 59-kDa protein synthesis initiation factor from wheat germ is functionally similar to the 80-kDa initiation factor 4B from mammalian cells. J Biol Chem. 1989 May 25;264(15):8491–8494. [PubMed] [Google Scholar]
  10. Browning K. S., Maia D. M., Lax S. R., Ravel J. M. Identification of a new protein synthesis initiation factor from wheat germ. J Biol Chem. 1987 Jan 15;262(2):538–541. [PubMed] [Google Scholar]
  11. Cottrelle P., Cool M., Thuriaux P., Price V. L., Thiele D., Buhler J. M., Fromageot P. Either one of the two yeast EF-1 alpha genes is required for cell viability. Curr Genet. 1985;9(8):693–697. doi: 10.1007/BF00449823. [DOI] [PubMed] [Google Scholar]
  12. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  13. Goyer C., Altmann M., Lee H. S., Blanc A., Deshmukh M., Woolford J. L., Jr, Trachsel H., Sonenberg N. TIF4631 and TIF4632: two yeast genes encoding the high-molecular-weight subunits of the cap-binding protein complex (eukaryotic initiation factor 4F) contain an RNA recognition motif-like sequence and carry out an essential function. Mol Cell Biol. 1993 Aug;13(8):4860–4874. doi: 10.1128/mcb.13.8.4860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gulyas K. D., Donahue T. F. SSL2, a suppressor of a stem-loop mutation in the HIS4 leader encodes the yeast homolog of human ERCC-3. Cell. 1992 Jun 12;69(6):1031–1042. doi: 10.1016/0092-8674(92)90621-i. [DOI] [PubMed] [Google Scholar]
  15. Hershey J. W. Overview: phosphorylation and translation control. Enzyme. 1990;44(1-4):17–27. doi: 10.1159/000468744. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Jaramillo M., Browning K., Dever T. E., Blum S., Trachsel H., Merrick W. C., Ravel J. M., Sonenberg N. Translation initiation factors that function as RNA helicases from mammals, plants and yeast. Biochim Biophys Acta. 1990 Aug 27;1050(1-3):134–139. doi: 10.1016/0167-4781(90)90154-t. [DOI] [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. Lawson T. G., Lee K. A., Maimone M. M., Abramson R. D., Dever T. E., Merrick W. C., Thach R. E. Dissociation of double-stranded polynucleotide helical structures by eukaryotic initiation factors, as revealed by a novel assay. Biochemistry. 1989 May 30;28(11):4729–4734. doi: 10.1021/bi00437a033. [DOI] [PubMed] [Google Scholar]
  21. Linder P., Lasko P. F., Ashburner M., Leroy P., Nielsen P. J., Nishi K., Schnier J., Slonimski P. P. Birth of the D-E-A-D box. Nature. 1989 Jan 12;337(6203):121–122. doi: 10.1038/337121a0. [DOI] [PubMed] [Google Scholar]
  22. Linder P. Molecular biology of translation in yeast. Antonie Van Leeuwenhoek. 1992 Aug;62(1-2):47–62. doi: 10.1007/BF00584462. [DOI] [PubMed] [Google Scholar]
  23. Linder P., Slonimski P. P. An essential yeast protein, encoded by duplicated genes TIF1 and TIF2 and homologous to the mammalian translation initiation factor eIF-4A, can suppress a mitochondrial missense mutation. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2286–2290. doi: 10.1073/pnas.86.7.2286. [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. Milburn S. C., Pelletier J., Sonenberg N., Hershey J. W. Identification of the 80-kDa protein that crosslinks to the cap structure of eukaryotic mRNAs as initiation factor eIF-4B. Arch Biochem Biophys. 1988 Jul;264(1):348–350. doi: 10.1016/0003-9861(88)90604-2. [DOI] [PubMed] [Google Scholar]
  26. Nielsen P. J., McMaster G. K., Trachsel H. Cloning of eukaryotic protein synthesis initiation factor genes: isolation and characterization of cDNA clones encoding factor eIF-4A. Nucleic Acids Res. 1985 Oct 11;13(19):6867–6880. doi: 10.1093/nar/13.19.6867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nielsen P. J., Trachsel H. The mouse protein synthesis initiation factor 4A gene family includes two related functional genes which are differentially expressed. EMBO J. 1988 Jul;7(7):2097–2105. doi: 10.1002/j.1460-2075.1988.tb03049.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Owttrim G. W., Hofmann S., Kuhlemeier C. Divergent genes for translation initiation factor eIF-4A are coordinately expressed in tobacco. Nucleic Acids Res. 1991 Oct 25;19(20):5491–5496. doi: 10.1093/nar/19.20.5491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Roemer T., Bussey H. Yeast beta-glucan synthesis: KRE6 encodes a predicted type II membrane protein required for glucan synthesis in vivo and for glucan synthase activity in vitro. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11295–11299. doi: 10.1073/pnas.88.24.11295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Schmid S. R., Linder P. Translation initiation factor 4A from Saccharomyces cerevisiae: analysis of residues conserved in the D-E-A-D family of RNA helicases. Mol Cell Biol. 1991 Jul;11(7):3463–3471. doi: 10.1128/mcb.11.7.3463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schnier J., Schwelberger H. G., Smit-McBride Z., Kang H. A., Hershey J. W. Translation initiation factor 5A and its hypusine modification are essential for cell viability in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1991 Jun;11(6):3105–3114. doi: 10.1128/mcb.11.6.3105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Stotz A., Linder P. The ADE2 gene from Saccharomyces cerevisiae: sequence and new vectors. Gene. 1990 Oct 30;95(1):91–98. doi: 10.1016/0378-1119(90)90418-q. [DOI] [PubMed] [Google Scholar]
  36. Teem J. L., Abovich N., Kaufer N. F., Schwindinger W. F., Warner J. R., Levy A., Woolford J., Leer R. J., van Raamsdonk-Duin M. M., Mager W. H. A comparison of yeast ribosomal protein gene DNA sequences. Nucleic Acids Res. 1984 Nov 26;12(22):8295–8312. doi: 10.1093/nar/12.22.8295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Waterston R., Martin C., Craxton M., Huynh C., Coulson A., Hillier L., Durbin R., Green P., Shownkeen R., Halloran N. A survey of expressed genes in Caenorhabditis elegans. Nat Genet. 1992 May;1(2):114–123. doi: 10.1038/ng0592-114. [DOI] [PubMed] [Google Scholar]
  38. Zaret K. S., Sherman F. DNA sequence required for efficient transcription termination in yeast. Cell. 1982 Mar;28(3):563–573. doi: 10.1016/0092-8674(82)90211-2. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES