Abstract
Eukaryotic mRNA molecules have a 5′ cap structure that is recognized by the cap-binding component of translation initiation factor eIF-4F during protein synthesis. In the budding yeast Saccharomyces cerevisiae this cap-binding protein is encoded by the CDC33 gene. We report here that decreased global translation initiation in cdc33 mutant cells has virtually no effect on the translation of mRNA from the SSA1 -lacZ chimeric gene, comprised of yeast SSA1 hsp70 gene transcription and translation initiation sequences fused in-frame to the bacterial lacZ gene. When global translation initiation was limited in cdc33 mutant cells, Ssal-LacZ polypeptide synthesis was increased relative to total protein synthesis, and the β-galactosidase activity of the SsaI-LacZ fusion protein was induced to wild-type levels. The normal rate of Ssal-LacZ polypeptide synthesis in mutant cells was maintained by normal levels of SSA1 -lacZ mRNA. Furthermore, in cdc33 mutant cells, the size of polysomes containing SSA1-lacZ mRNA was unaffected, while polysomes containing other specific mRNAs were smaller. Efficient Ssal-LacZ polypeptide synthesis was also seen during eIF-4F limitation produced by disruption of the TIF4631 gene, encoding the large eIF-4F subunit. All of these findings indicate efficient SSA1-lacZ mRNA usage under conditions of globally impaired translation initiation due to eIF-4F limitation.
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References
Altmann M, Trachsel H (1989) Altered mRNA cap recognition activity of initiation factor 4E in the yeast cell cycle division mutant cdc33. Nucleic Acids Res 17:5923–5931
Altmann M, Handschin C, Trachsel H (1987) mRNA cap-binding protein: cloning of the gene encoding protein synthesis initiation factor eIF-4E from Saccharomyces cerevisiae. Mol Cell Biol 7:998–1003
Altmann M, Sonenberg N, Trachsel H (1989) Translation in Saccharomyces cerevisiae: initiation factor 4E-dependent cell-free system. Mol Cell Biol 9:4467–4472
Barnes CA, Johnston GC, Singer RA (1991) Expression of lacZ gene fusions affects downstream transcription in yeast. Gene 104:47–54
Barnes CA, Singer RA, Johnston GC (1993) Yeast prt1 mutations alter heat-shock gene expression through transcript fragmentation. EMBO J 12:3323–3332
Boorstein WR, Craig EA (1990) Transcriptional regulation of SSA3, an HSP70 gene from Saccharomyces cerevisiae. Mol Cell Biol 10:3262–3267
Boorstein WR, Ziegelhoffer T, Craig EA (1994) Molecular evolution of the HSP70 multigene family. J Mol Evol 38:1–17
Brenner C, Nakayama N, Goebl M, Tanaka K, Toh-e A, Matsumoto K (1988) CDC33 encodes mRNA cap-binding protein eIF-4E of Saccharomyces cerevisiae. Mol Cell Biol 8:3556–3559
Cigan AM, Foiani M, Hannig EM, Hinnebusch AG (1991) Complex formation by positive and negative translational regulators of GCN4. Mol Cell Biol 11:3217–3228
De Benedetti A, Joshi-Barve S, Rinker-Schaeffer C, Rhoads RE (1991) Expression of antisense RNA against initiation factor eIF-4E mRNA in HeLa cells results in lengthened cell division times, diminished translation rates, and reduced levels of both eIF-4E and the p220 component of eIF-4E. Mol Cell Biol 11:5435–5445
Duncan R, Hershey JWB (1984) Heat shock-induced translational alterations in HeLa cells: initiation factor modifications and the inhibition of translation. J Biol Chem 259:11882–11889
Duncan R, Milburn SC, Hershey JWB (1987) Regulated phosphorylation and low abundance of HeLa cell initiation factor cIF4F suggest a role in translational control: heat shock effects on eIF-4F. J Biol Chem 262:380–388
Ellwood MS, Craig EA (1984) Differential regulation of the 70 K heat shock gene and related genes in Saccharomyces cerevisiae. Mol Cell Biol 4:1454–1459
Foiani M, Cigan AM, Paddon CJ, Harashima S, Hinnebusch AG (1991) GCD2, a translational repressor of the GCN4 gene, has a general function in the initiation of protein synthesis in Saccharomyces cerevisiae. Mol Cell Biol 11:3203–3216
Goyer C, Altmann M, Trachsel H, Sonenberg N (1989) Identification and characterization of cap-binding proteins from yeast. J Biol Chem 264:7603–7610
Goyer C, Altmann M, Lee HS, Blanc A, Deshmukh M, Woolford JL Jr, Trachsel H, Sonenberg N (1993) 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. Mot Cell Biol 13:4860–4874
Hartwell LH, McLaughlin CS (1969) A mutant of yeast apparently defective in the initiation of protein synthesis. Proc Natl Acad Sci USA 62:468–474
Iida H, Yahara I (1984a) Specific early-G1 blocks accompanied with stringent response in Saccharomyces cerevisiae lead to growth arrest in resting state similar to the Go of higher eucaryotes. J Cell Biol 98:1185–1193
Iida H, Yahara I (1984b) Durable synthesis of high molecular weight heat shock proteins in Go cells of the yeast and other eucaryotes. J Cell Biol 99:199–207
Joshi-Barve S, De Benedetti A, Rhoads RE (1992) Preferential translation of heat shock mRNAs in HeLa cells deficient in protein synthesis initiation factors eIF-4E and eIF-4γ. J Biol Chem 267:21038–21043
Klemenz R, Hultmark D, Gehring WJ (1985) Selective translation of heat-shock mRNA in Drosophila melanogaster depends on sequence information in the leader. EMBO J 4:2053–2060
Lamphear BJ, Panniers R (1990) Cap binding protein complex that restores protein synthesis in heat-shocked Ehrlich cell lysates contains highly phosphorylated eIF-4E. J Biol Chem 265:5333–5336
Lamphear BJ, Panniers R (1991) Heat shock impairs the interaction of cap-binding protein complex with 5′ mRNA cap. J Biol Chem 266:2789–2794
Lanker S, Müller PP, Altmann M, Goyer C, Sonenberg N, Trachsel H (1992) Interactions of the eIF-4F subunits in the yeast Saccharomyces cerevisiae. J Biol Chem 267:21167–21171
Lindquist S, Petersen R (1990) Selective translation and degradation of heat-shock messenger RNAs in Drosophila. Enzyme 44:147–166
Maroto FG, Sierra JM (1988) Translational control in heatshocked Drosophila embryos. J Biol Chem 263:15720–15725
McAlister L, Strausberg S, Kulaga A, Finkelstein DB (1979) Altered patterns of protein synthesis induced by heat shock of yeast. Carr Genet 1:63–74
McGarry TJ, Lindquist S (1985) The preferential translation of Drosophila hsp70 mRNA requires sequences in the untranslated leader. Cell 42:903–911
Moreland RB, Nam HG, Hereford LM, Fried HM (1985) Identification of a nuclear localization signal of a yeast ribosomal protein. Proc Natl Acad Sci USA 82:6561–6565
Pelletier J, Sonenberg N (1985) Insertion mutagenesis to increase secondary structure within the 5′ noncoding region of a eukaryotic mRNA reduces translational efficiency. Cell 40:515–526
Pelletier J, Sonenberg N (1988) Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA. Nature 334:320–325
Rhoads RE (1988) Cap recognition and the entry of mRNA into the protein synthesis initiation cycle. Trends Biochem Sci 13:52–56
Sachs AB, Davis RW (1989) The poly(A) binding protein is required for poly(A) shortening and 60S ribosomal subunit-dependent translation initiation. Cell 58:857–867
Sachs AB, Deardorff JA (1992) Translation initiation requires the PAB-dependent poly(A) ribonuclease in yeast. Cell 70:961–973
Slater MR, Craig EA (1987) Transcriptional regulation of an hsp70 heat shock gene in the yeast Saccharomyces cerevisiae. Mol Cell Biol 7:1906–1916
Slater MR, Craig EA (1989) The SSAI and SSA2 genes of the yeast Saccharomyces cerevisiae. Nucleic Acids Res 17:805–806
Sonenberg N (1988) Cap-binding proteins of eukaryotic messenger RNA: functions in initiation and control of translation. Prog Nucleic Acids Res Mol Biol 35:173–207
Thach RE (1992) Cap recap: the involvement of cIF-4F in regulating gene expression. Cell 68:177–180
Werner-Washburne M, Stone DE, Craig EA (1987) Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae. Mol Cell Biol 7:2568–2577
Yoon H, Donahue TF (1992) The suil suppressor locus in Saccharomyces cerevisiae encodes a translation factor that functions during tRNAi Met recognition of the start codon. Mol Cell Biol 12:248–260
Yoon H, Miller SP, Pabich EK, Donahue TF (1992) SSLI, a suppressor of a HIS4 5′-UTR stem-loop mutation, is essential for translation initiation and affects UV resistance in yeast. Genes Dev 6:2463–2477
Zapata JM, Maroto FG, Sierra JM (1991) Inactivation of mRNA cap-binding protein complex in Drosophila melanogaster embryos under heat shock. J Biol Chem 266:16007–16014
Zhong T, Arndt KT (1993) The yeast SIS1 protein, a DnaJ homolog, is required for the initiation of translation. Cell 73:1175–1186
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Communicated by D. Y. Thomas
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Barnes, C.A., MacKenzie, M.M., Johnston, G.C. et al. Efficient translation of an SSA1-derived heat-shock mRNA in yeast cells limited for cap-binding protein and eIF-4F. Molec. Gen. Genet. 246, 619–627 (1995). https://doi.org/10.1007/BF00298969
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DOI: https://doi.org/10.1007/BF00298969