Skip to main content
PMC home page
Genetics logoLink to Genetics
. 1994 Jan;136(1):75–86. doi: 10.1093/genetics/136.1.75

Translational Maintenance of Frame: Mutants of Saccharomyces Cerevisiae with Altered -1 Ribosomal Frameshifting Efficiences

J D Dinman 1, R B Wickner 1
PMCID: PMC1205794  PMID: 8138178

Abstract

A special site on the (+) strand of the L-A dsRNA virus induces about 2% of ribosomes translating the gag open reading frame to execute a -1 frameshift and thus produce the viral gag-pol fusion protein. Using constructs in which a -1 ribosomal frameshift at this site was necessary for expression of lacZ we isolated chromosomal mutants in which the efficiency of frameshifting was increased. These mutants comprise eight genes, named mof (maintenance of frame). The mof1-1, mof2-1, mof4-1, mof5-1 and mof6-1 strains cannot maintain M(1) dsRNA at 30°, but, paradoxically, do not lose L-A. The mof2-1, mof5-1 and mof6-1 strains are temperature sensitive for growth at 37°, and all three show striking cell cycle phenotypes. The mof2-1 strains arrest with mother and daughter cells almost equal in size, mof5-1 arrests with multiple buds and mof6-1 arrests as single large unbudded cells. mof2-1 and mof5-1 strains are also Pet(-). The mof mutations show differential effects on various frameshifting signals.

Full Text

The Full Text of this article is available as a PDF (2.6 MB).

Selected References

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

  1. Atkins J. F., Weiss R. B., Thompson S., Gesteland R. F. Towards a genetic dissection of the basis of triplet decoding, and its natural subversion: programmed reading frame shifts and hops. Annu Rev Genet. 1991;25:201–228. doi: 10.1146/annurev.ge.25.120191.001221. [DOI] [PubMed] [Google Scholar]
  2. Ball S. G., Tirtiaux C., Wickner R. B. Genetic Control of L-a and L-(Bc) Dsrna Copy Number in Killer Systems of SACCHAROMYCES CEREVISIAE. Genetics. 1984 Jun;107(2):199–217. doi: 10.1093/genetics/107.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Belcourt M. F., Farabaugh P. J. Ribosomal frameshifting in the yeast retrotransposon Ty: tRNAs induce slippage on a 7 nucleotide minimal site. Cell. 1990 Jul 27;62(2):339–352. doi: 10.1016/0092-8674(90)90371-K. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brierley I., Digard P., Inglis S. C. Characterization of an efficient coronavirus ribosomal frameshifting signal: requirement for an RNA pseudoknot. Cell. 1989 May 19;57(4):537–547. doi: 10.1016/0092-8674(89)90124-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brierley I., Jenner A. J., Inglis S. C. Mutational analysis of the "slippery-sequence" component of a coronavirus ribosomal frameshifting signal. J Mol Biol. 1992 Sep 20;227(2):463–479. doi: 10.1016/0022-2836(92)90901-U. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cox B. S., Tuite M. F., McLaughlin C. S. The psi factor of yeast: a problem in inheritance. Yeast. 1988 Sep;4(3):159–178. doi: 10.1002/yea.320040302. [DOI] [PubMed] [Google Scholar]
  8. Cummins C. M., Gaber R. F., Culbertson M. R., Mann R., Fink G. R. Frameshift suppression in Saccharomyces cerevisiae. III. Isolation and genetic properties of group III suppressors. Genetics. 1980 Aug;95(4):855–879. doi: 10.1093/genetics/95.4.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dihanich M. E., Najarian D., Clark R., Gillman E. C., Martin N. C., Hopper A. K. Isolation and characterization of MOD5, a gene required for isopentenylation of cytoplasmic and mitochondrial tRNAs of Saccharomyces cerevisiae. Mol Cell Biol. 1987 Jan;7(1):177–184. doi: 10.1128/mcb.7.1.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dinman J. D., Icho T., Wickner R. B. A -1 ribosomal frameshift in a double-stranded RNA virus of yeast forms a gag-pol fusion protein. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):174–178. doi: 10.1073/pnas.88.1.174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dinman J. D., Wickner R. B. Ribosomal frameshifting efficiency and gag/gag-pol ratio are critical for yeast M1 double-stranded RNA virus propagation. J Virol. 1992 Jun;66(6):3669–3676. doi: 10.1128/jvi.66.6.3669-3676.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eng W. K., Pandit S. D., Sternglanz R. Mapping of the active site tyrosine of eukaryotic DNA topoisomerase I. J Biol Chem. 1989 Aug 15;264(23):13373–13376. [PubMed] [Google Scholar]
  13. Esteban R., Fujimura T., Wickner R. B. Site-specific binding of viral plus single-stranded RNA to replicase-containing open virus-like particles of yeast. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4411–4415. doi: 10.1073/pnas.85.12.4411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Esteban R., Wickner R. B. A deletion mutant of L-A double-stranded RNA replicates like M1 double-stranded RNA. J Virol. 1988 Apr;62(4):1278–1285. doi: 10.1128/jvi.62.4.1278-1285.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Esteban R., Wickner R. B. Three different M1 RNA-containing viruslike particle types in Saccharomyces cerevisiae: in vitro M1 double-stranded RNA synthesis. Mol Cell Biol. 1986 May;6(5):1552–1561. doi: 10.1128/mcb.6.5.1552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  17. Fried H. M., Fink G. R. Electron microscopic heteroduplex analysis of "killer" double-stranded RNA species from yeast. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4224–4228. doi: 10.1073/pnas.75.9.4224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fujimura T., Ribas J. C., Makhov A. M., Wickner R. B. Pol of gag-pol fusion protein required for encapsidation of viral RNA of yeast L-A virus. Nature. 1992 Oct 22;359(6397):746–749. doi: 10.1038/359746a0. [DOI] [PubMed] [Google Scholar]
  19. Fujimura T., Wickner R. B. Gene overlap results in a viral protein having an RNA binding domain and a major coat protein domain. Cell. 1988 Nov 18;55(4):663–671. doi: 10.1016/0092-8674(88)90225-5. [DOI] [PubMed] [Google Scholar]
  20. Fujimura T., Wickner R. B. Interaction of two cis sites with the RNA replicase of the yeast L-A virus. J Biol Chem. 1992 Feb 5;267(4):2708–2713. [PubMed] [Google Scholar]
  21. Guarente L. Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol. 1983;101:181–191. doi: 10.1016/0076-6879(83)01013-7. [DOI] [PubMed] [Google Scholar]
  22. Hatfield D. L., Levin J. G., Rein A., Oroszlan S. Translational suppression in retroviral gene expression. Adv Virus Res. 1992;41:193–239. doi: 10.1016/S0065-3527(08)60037-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hopper J. E., Bostian K. A., Rowe L. B., Tipper D. J. Translation of the L-species dsRNA genome of the killer-associated virus-like particles of Saccharomyces cerevisiae. J Biol Chem. 1977 Dec 25;252(24):9010–9017. [PubMed] [Google Scholar]
  24. 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]
  25. Jacks T., Madhani H. D., Masiarz F. R., Varmus H. E. Signals for ribosomal frameshifting in the Rous sarcoma virus gag-pol region. Cell. 1988 Nov 4;55(3):447–458. doi: 10.1016/0092-8674(88)90031-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Jacks T. Translational suppression in gene expression in retroviruses and retrotransposons. Curr Top Microbiol Immunol. 1990;157:93–124. doi: 10.1007/978-3-642-75218-6_4. [DOI] [PubMed] [Google Scholar]
  27. Lawrence C. W. Classical mutagenesis techniques. Methods Enzymol. 1991;194:273–281. doi: 10.1016/0076-6879(91)94021-4. [DOI] [PubMed] [Google Scholar]
  28. Ribas J. C., Wickner R. B. RNA-dependent RNA polymerase consensus sequence of the L-A double-stranded RNA virus: definition of essential domains. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2185–2189. doi: 10.1073/pnas.89.6.2185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ridley S. P., Sommer S. S., Wickner R. B. Superkiller mutations in Saccharomyces cerevisiae suppress exclusion of M2 double-stranded RNA by L-A-HN and confer cold sensitivity in the presence of M and L-A-HN. Mol Cell Biol. 1984 Apr;4(4):761–770. doi: 10.1128/mcb.4.4.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tu C., Tzeng T. H., Bruenn J. A. Ribosomal movement impeded at a pseudoknot required for frameshifting. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8636–8640. doi: 10.1073/pnas.89.18.8636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tuite M. F., Mundy C. R., Cox B. S. Agents that cause a high frequency of genetic change from [psi+] to [psi-] in Saccharomyces cerevisiae. Genetics. 1981 Aug;98(4):691–711. doi: 10.1093/genetics/98.4.691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Uemura H., Wickner R. B. Suppression of chromosomal mutations affecting M1 virus replication in Saccharomyces cerevisiae by a variant of a viral RNA segment (L-A) that encodes coat protein. Mol Cell Biol. 1988 Feb;8(2):938–944. doi: 10.1128/mcb.8.2.938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Valle R. P., Wickner R. B. Elimination of L-A double-stranded RNA virus of Saccharomyces cerevisiae by expression of gag and gag-pol from an L-A cDNA clone. J Virol. 1993 May;67(5):2764–2771. doi: 10.1128/jvi.67.5.2764-2771.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wakem L. P., Sherman F. Isolation and characterization of omnipotent suppressors in the yeast Saccharomyces cerevisiae. Genetics. 1990 Mar;124(3):515–522. doi: 10.1093/genetics/124.3.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wickner R. B., Leibowitz M. J. Two chromosomal genes required for killing expression in killer strains of Saccharomyces cerevisiae. Genetics. 1976 Mar 25;82(3):429–442. doi: 10.1093/genetics/82.3.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Xu H., Boeke J. D. Host genes that influence transposition in yeast: the abundance of a rare tRNA regulates Ty1 transposition frequency. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8360–8364. doi: 10.1073/pnas.87.21.8360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zhu Y. S., Kane J., Zhang X. Y., Zhang M., Tipper D. J. Role of the gamma component of preprotoxin in expression of the yeast K1 killer phenotype. Yeast. 1993 Mar;9(3):251–266. doi: 10.1002/yea.320090305. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES