• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Sep 1, 1995; 14(17): 4365–4373.
PMCID: PMC394521

The products of the SUP45 (eRF1) and SUP35 genes interact to mediate translation termination in Saccharomyces cerevisiae.


The product of the yeast SUP45 gene (Sup45p) is highly homologous to the Xenopus eukaryote release factor 1 (eRF1), which has release factor activity in vitro. We show, using the two-hybrid system, that in Saccharomyces cerevisiae Sup45p and the product of the SUP35 gene (Sup35p) interact in vivo. The ability of Sup45p C-terminally tagged with (His)6 to specifically precipitate Sup35p from a cell lysate was used to confirm this interaction in vitro. Although overexpression of either the SUP45 or SUP35 genes alone did not reduce the efficiency of codon-specific tRNA nonsense suppression, the simultaneous overexpression of both the SUP35 and SUP45 genes in nonsense suppressor tRNA-containing strains produced an antisuppressor phenotype. These data are consistent with Sup35p and Sup45p forming a complex with release factor properties. Furthermore, overexpression of either Xenopus or human eRF1 (SUP45) genes also resulted in anti-suppression only if that strain was also overexpressing the yeast SUP35 gene. Antisuppression is a characteristic phenotype associated with overexpression of both prokaryote and mitochondrial release factors. We propose that Sup45p and Sup35p interact to form a release factor complex in yeast and that Sup35p, which has GTP binding sequence motifs in its C-terminal domain, provides the GTP hydrolytic activity which is a demonstrated requirement of the eukaryote translation termination reaction.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bartel P, Chien CT, Sternglanz R, Fields S. Elimination of false positives that arise in using the two-hybrid system. Biotechniques. 1993 Jun;14(6):920–924. [PubMed]
  • Becker DM, Guarente L. High-efficiency transformation of yeast by electroporation. Methods Enzymol. 1991;194:182–187. [PubMed]
  • Breeden L, Nasmyth K. Regulation of the yeast HO gene. Cold Spring Harb Symp Quant Biol. 1985;50:643–650. [PubMed]
  • Caskey CT, Tompkins R, Scolnick E, Caryk T, Nirenberg M. Sequential translation of trinucleotide codons for the initiation and termination of protein synthesis. Science. 1968 Oct 4;162(3849):135–138. [PubMed]
  • Chernoff YO, Derkach IL, Inge-Vechtomov SG. Multicopy SUP35 gene induces de-novo appearance of psi-like factors in the yeast Saccharomyces cerevisiae. Curr Genet. 1993 Sep;24(3):268–270. [PubMed]
  • Chien CT, Bartel PL, 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. [PMC free article] [PubMed]
  • Christianson TW, Sikorski RS, Dante M, Shero JH, Hieter P. Multifunctional yeast high-copy-number shuttle vectors. Gene. 1992 Jan 2;110(1):119–122. [PubMed]
  • Cox B. Cytoplasmic inheritance. Prion-like factors in yeast. Curr Biol. 1994 Aug 1;4(8):744–748. [PubMed]
  • Didichenko SA, Ter-Avanesyan MD, Smirnov VN. Ribosome-bound EF-1 alpha-like protein of yeast Saccharomyces cerevisiae. Eur J Biochem. 1991 Jun 15;198(3):705–711. [PubMed]
  • Doel SM, McCready SJ, Nierras CR, Cox BS. The dominant PNM2- mutation which eliminates the psi factor of Saccharomyces cerevisiae is the result of a missense mutation in the SUP35 gene. Genetics. 1994 Jul;137(3):659–670. [PMC free article] [PubMed]
  • Dowell SJ, Tsang JS, Mellor J. The centromere and promoter factor 1 of yeast contains a dimerisation domain located carboxy-terminal to the bHLH domain. Nucleic Acids Res. 1992 Aug 25;20(16):4229–4236. [PMC free article] [PubMed]
  • Finkelstein DB, Strausberg S. Heat shock-regulated production of Escherichia coli beta-galactosidase in Saccharomyces cerevisiae. Mol Cell Biol. 1983 Sep;3(9):1625–1633. [PMC free article] [PubMed]
  • Fields S, Song O. A novel genetic system to detect protein-protein interactions. Nature. 1989 Jul 20;340(6230):245–246. [PubMed]
  • Frolova LYu, Dalphin ME, Justesen J, Powell RJ, Drugeon G, McCaughan KK, Kisselev LL, Tate WP, Haenni AL. Mammalian polypeptide chain release factor and tryptophanyl-tRNA synthetase are distinct proteins. EMBO J. 1993 Oct;12(10):4013–4019. [PMC free article] [PubMed]
  • Frolova L, Le Goff X, Rasmussen HH, Cheperegin S, Drugeon G, Kress M, Arman I, Haenni AL, Celis JE, Philippe M, et al. A highly conserved eukaryotic protein family possessing properties of polypeptide chain release factor. Nature. 1994 Dec 15;372(6507):701–703. [PubMed]
  • Goldstein JL, Beaudet AL, Caskey CT. Peptide chain termination with mammalian release factor. Proc Natl Acad Sci U S A. 1970 Sep;67(1):99–106. [PMC free article] [PubMed]
  • Grenett HE, Bounelis P, Fuller GM. Identification of a human cDNA with high homology to yeast omnipotent suppressor 45. Gene. 1992 Jan 15;110(2):239–243. [PubMed]
  • Grentzmann G, Brechemier-Baey D, Heurgue V, Mora L, Buckingham RH. Localization and characterization of the gene encoding release factor RF3 in Escherichia coli. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):5848–5852. [PMC free article] [PubMed]
  • Hawthorne DC, Leupold U. Suppressors in yeast. Curr Top Microbiol Immunol. 1974;64(0):1–47. [PubMed]
  • Himmelfarb HJ, Maicas E, Friesen JD. Isolation of the SUP45 omnipotent suppressor gene of Saccharomyces cerevisiae and characterization of its gene product. Mol Cell Biol. 1985 Apr;5(4):816–822. [PMC free article] [PubMed]
  • Hirashima A, Kaji A. Factor-dependent release of ribosomes from messenger RNA. Requirement for two heat-stable factors. J Mol Biol. 1972 Mar 14;65(1):43–58. [PubMed]
  • Hoshino S, Miyazawa H, Enomoto T, Hanaoka F, Kikuchi Y, Kikuchi A, Ui M. A human homologue of the yeast GST1 gene codes for a GTP-binding protein and is expressed in a proliferation-dependent manner in mammalian cells. EMBO J. 1989 Dec 1;8(12):3807–3814. [PMC free article] [PubMed]
  • Jones JS, Prakash L. Yeast Saccharomyces cerevisiae selectable markers in pUC18 polylinkers. Yeast. 1990 Sep-Oct;6(5):363–366. [PubMed]
  • Klein H, Capecchi MR. Polypetide chain termination. Purification of the release factors, R1 and R2, from Escherichia coli. J Biol Chem. 1971 Feb 25;246(4):1055–1061. [PubMed]
  • Konecki DS, Aune KC, Tate W, Caskey CT. Characterization of reticulocyte release factor. J Biol Chem. 1977 Jul 10;252(13):4514–4520. [PubMed]
  • Kushnirov VV, Ter-Avanesyan MD, Telckov MV, Surguchov AP, Smirnov VN, Inge-Vechtomov SG. Nucleotide sequence of the SUP2 (SUP35) gene of Saccharomyces cerevisiae. Gene. 1988 Jun 15;66(1):45–54. [PubMed]
  • Kushnirov VV, Ter-Avanesyan MD, Didichenko SA, Smirnov VN, Chernoff YO, Derkach IL, Novikova ON, Inge-Vechtomov SG, Neistat MA, Tolstorukov II. Divergence and conservation of SUP2 (SUP35) gene of yeast Pichia pinus and Saccharomyces cerevisiae. Yeast. 1990 Nov-Dec;6(6):461–472. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lee CC, Craigen WJ, Muzny DM, Harlow E, Caskey CT. Cloning and expression of a mammalian peptide chain release factor with sequence similarity to tryptophanyl-tRNA synthetases. Proc Natl Acad Sci U S A. 1990 May;87(9):3508–3512. [PMC free article] [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Milman G, Goldstein J, Scolnick E, Caskey T. Peptide chain termination. 3. Stimulation of in vitro termination. Proc Natl Acad Sci U S A. 1969 May;63(1):183–190. [PMC free article] [PubMed]
  • Mikuni O, Ito K, Moffat J, Matsumura K, McCaughan K, Nobukuni T, Tate W, Nakamura Y. Identification of the prfC gene, which encodes peptide-chain-release factor 3 of Escherichia coli. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):5798–5802. [PMC free article] [PubMed]
  • Pel HJ, Maat C, Rep M, Grivell LA. The yeast nuclear gene MRF1 encodes a mitochondrial peptide chain release factor and cures several mitochondrial RNA splicing defects. Nucleic Acids Res. 1992 Dec 11;20(23):6339–6346. [PMC free article] [PubMed]
  • Pure GA, Robinson GW, Naumovski L, Friedberg EC. Partial suppression of an ochre mutation in Saccharomyces cerevisiae by multicopy plasmids containing a normal yeast tRNAGln gene. J Mol Biol. 1985 May 5;183(1):31–42. [PubMed]
  • Scolnick E, Tompkins R, Caskey T, Nirenberg M. Release factors differing in specificity for terminator codons. Proc Natl Acad Sci U S A. 1968 Oct;61(2):768–774. [PMC free article] [PubMed]
  • Sherman F. Getting started with yeast. Methods Enzymol. 1991;194:3–21. [PubMed]
  • Song JM, Liebman SW. Allosuppressors that enhance the efficiency of omnipotent suppressors in Saccharomyces cerevisiae. Genetics. 1987 Mar;115(3):451–460. [PMC free article] [PubMed]
  • Stansfield I, Grant GM, Akhmaloka, Tuite MF. Ribosomal association of the yeast SAL4 (SUP45) gene product: implications for its role in translation fidelity and termination. Mol Microbiol. 1992 Dec;6(23):3469–3478. [PubMed]
  • Stansfield I, Akhmaloka, Tuite MF. A mutant allele of the SUP45 (SAL4) gene of Saccharomyces cerevisiae shows temperature-dependent allosuppressor and omnipotent suppressor phenotypes. Curr Genet. 1995 Apr;27(5):417–426. [PubMed]
  • Tassan JP, Le Guellec K, Kress M, Faure M, Camonis J, Jacquet M, Philippe M. In Xenopus laevis, the product of a developmentally regulated mRNA is structurally and functionally homologous to a Saccharomyces cerevisiae protein involved in translation fidelity. Mol Cell Biol. 1993 May;13(5):2815–2821. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Tuite MF. Genetics. Psi no more for yeast prions. Nature. 1994 Aug 4;370(6488):327–328. [PubMed]
  • Tuite MF, McLaughlin CS. Endogenous read-through of a UGA termination codon in a Saccharomyces cerevisiae cell-free system: evidence for involvement of both a mitochondrial and a nuclear tRNA. Mol Cell Biol. 1982 May;2(5):490–497. [PMC free article] [PubMed]
  • Tuite MF, Stansfield I. Translation. Knowing when to stop. Nature. 1994 Dec 15;372(6507):614–615. [PubMed]
  • Vincent A, Newnam G, Liebman SW. The yeast translational allosuppressor, SAL6: a new member of the PP1-like phosphatase family with a long serine-rich N-terminal extension. Genetics. 1994 Nov;138(3):597–608. [PMC free article] [PubMed]
  • Weiss RB, Murphy JP, Gallant JA. Genetic screen for cloned release factor genes. J Bacteriol. 1984 Apr;158(1):362–364. [PMC free article] [PubMed]
  • Weiss WA, Friedberg EC. Normal yeast tRNA(CAGGln) can suppress amber codons and is encoded by an essential gene. J Mol Biol. 1986 Dec 20;192(4):725–735. [PubMed]
  • Wickner RB. [URE3] as an altered URE2 protein: evidence for a prion analog in Saccharomyces cerevisiae. Science. 1994 Apr 22;264(5158):566–569. [PubMed]
  • Wilson PG, Culbertson MR. SUF12 suppressor protein of yeast. A fusion protein related to the EF-1 family of elongation factors. J Mol Biol. 1988 Feb 20;199(4):559–573. [PubMed]
  • Zhouravleva G, Frolova L, Le Goff X, Le Guellec R, Inge-Vechtomov S, Kisselev L, Philippe M. Termination of translation in eukaryotes is governed by two interacting polypeptide chain release factors, eRF1 and eRF3. EMBO J. 1995 Aug 15;14(16):4065–4072. [PMC free article] [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...