Skip to main content
PMC home page
Genetics logoLink to Genetics
. 1994 Nov;138(3):597–607. doi: 10.1093/genetics/138.3.597

The Yeast Translational Allosuppressor, Sal6: A New Member of the Pp1-like Phosphatase Family with a Long Serine-Rich N-Terminal Extension

A Vincent 1, G Newnam 1, S W Liebman 1
PMCID: PMC1206211  PMID: 7851758

Abstract

The allosuppressor mutation, sal6-1, enhances the efficiency of all tested translational suppressors, including codon-specific tRNA suppressors as well as codon-nonspecific omnipotent suppressors. The SAL6 gene has now been cloned by complementation of the increased suppression efficiency and cold sensitivity caused by sal6-1 in the presence of the omnipotent suppressor sup45. Physical analysis maps SAL6 to chromosome XVI between TPK2 and spt14. The SAL6 gene encodes a very basic 549-amino acid protein whose C-terminal catalytic region of 265 residues is 63% identical to serine/threonine PP1 phosphatases, and 66% identical to yeast PPZ1 and PPZ2 phosphatases. The unusual 235 residue N-terminal extension found in SAL6, like those in the PPZ proteins, is serine-rich. The sal6-1 mutation is a frameshift at amino acid position 271 which destroys the presumed phosphatase catalytic domain of the protein. Disruptions of the entire SAL6 gene are viable, cause a slight growth defect on glycerol medium, and produce allosuppressor phenotypes in suppressor strain backgrounds. The role of the serine-rich N terminus is unclear, since sal6 phenotypes are fully complemented by a SAL6 allele that contains an in-frame deletion of most of this region. High copy number plasmids containing wild-type SAL6 cause antisuppressor phenotypes in suppressor strains. These results suggest that the accuracy of protein synthesis is affected by the levels of phosphorylation of the target(s) of SAL6.

Full Text

The Full Text of this article is available as a PDF (5.0 MB).

Selected References

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

  1. All-Robyn J. A., Kelley-Geraghty D., Griffin E., Brown N., Liebman S. W. Isolation of omnipotent suppressors in an [eta+] yeast strain. Genetics. 1990 Mar;124(3):505–514. doi: 10.1093/genetics/124.3.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Arndt K. T., Styles C. A., Fink G. R. A suppressor of a HIS4 transcriptional defect encodes a protein with homology to the catalytic subunit of protein phosphatases. Cell. 1989 Feb 24;56(4):527–537. doi: 10.1016/0092-8674(89)90576-x. [DOI] [PubMed] [Google Scholar]
  4. Bennetzen J. L., Hall B. D. Codon selection in yeast. J Biol Chem. 1982 Mar 25;257(6):3026–3031. [PubMed] [Google Scholar]
  5. Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
  6. Chen M. X., Chen Y. H., Cohen P. T. PPQ, a novel protein phosphatase containing a Ser + Asn-rich amino-terminal domain, is involved in the regulation of protein synthesis. Eur J Biochem. 1993 Dec 1;218(2):689–699. doi: 10.1111/j.1432-1033.1993.tb18423.x. [DOI] [PubMed] [Google Scholar]
  7. Chen M. X., Chen Y. H., Cohen P. T. Polymerase chain reactions using Saccharomyces, Drosophila and human DNA predict a large family of protein serine/threonine phosphatases. FEBS Lett. 1992 Jul 13;306(1):54–58. doi: 10.1016/0014-5793(92)80836-6. [DOI] [PubMed] [Google Scholar]
  8. Chernoff Y. O., Vincent A., Liebman S. W. Mutations in eukaryotic 18S ribosomal RNA affect translational fidelity and resistance to aminoglycoside antibiotics. EMBO J. 1994 Feb 15;13(4):906–913. doi: 10.1002/j.1460-2075.1994.tb06334.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clotet J., Posas F., Casamayor A., Schaaff-Gerstenschläger I., Ariño J. The gene DIS2S1 is essential in Saccharomyces cerevisiae and is involved in glycogen phosphorylase activation. Curr Genet. 1991 May;19(5):339–342. doi: 10.1007/BF00309593. [DOI] [PubMed] [Google Scholar]
  10. Cohen P., Cohen P. T. Protein phosphatases come of age. J Biol Chem. 1989 Dec 25;264(36):21435–21438. [PubMed] [Google Scholar]
  11. Crouzet M., Izgu F., Grant C. M., Tuite M. F. The allosuppressor gene SAL4 encodes a protein important for maintaining translational fidelity in Saccharomyces cerevisiae. Curr Genet. 1988 Dec;14(6):537–543. doi: 10.1007/BF00434078. [DOI] [PubMed] [Google Scholar]
  12. Crouzet M., Tuite M. F. Genetic control of translational fidelity in yeast: molecular cloning and analysis of the allosuppressor gene SAL3. Mol Gen Genet. 1987 Dec;210(3):581–583. doi: 10.1007/BF00327216. [DOI] [PubMed] [Google Scholar]
  13. Cyert M. S., Kunisawa R., Kaim D., Thorner J. Yeast has homologs (CNA1 and CNA2 gene products) of mammalian calcineurin, a calmodulin-regulated phosphoprotein phosphatase. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7376–7380. doi: 10.1073/pnas.88.16.7376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Davidow L. S., Byers B. Enhanced gene conversion and postmeiotic segregation in pachytene-arrested Saccharomyces cerevisiae. Genetics. 1984 Feb;106(2):165–183. doi: 10.1093/genetics/106.2.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Elble R. A simple and efficient procedure for transformation of yeasts. Biotechniques. 1992 Jul;13(1):18–20. [PubMed] [Google Scholar]
  16. Gautier J., Solomon M. J., Booher R. N., Bazan J. F., Kirschner M. W. cdc25 is a specific tyrosine phosphatase that directly activates p34cdc2. Cell. 1991 Oct 4;67(1):197–211. doi: 10.1016/0092-8674(91)90583-k. [DOI] [PubMed] [Google Scholar]
  17. Grenett H. E., Bounelis P., Fuller G. M. Identification of a human cDNA with high homology to yeast omnipotent suppressor 45. Gene. 1992 Jan 15;110(2):239–243. doi: 10.1016/0378-1119(92)90655-9. [DOI] [PubMed] [Google Scholar]
  18. Hauber J., Stucka R., Krieg R., Feldmann H. Analysis of yeast chromosomal regions carrying members of the glutamate tRNA gene family: various transposable elements are associated with them. Nucleic Acids Res. 1988 Nov 25;16(22):10623–10634. doi: 10.1093/nar/16.22.10623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hershey J. W. Protein phosphorylation controls translation rates. J Biol Chem. 1989 Dec 15;264(35):20823–20826. [PubMed] [Google Scholar]
  20. Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
  21. Higgins D. G., Sharp P. M. Fast and sensitive multiple sequence alignments on a microcomputer. Comput Appl Biosci. 1989 Apr;5(2):151–153. doi: 10.1093/bioinformatics/5.2.151. [DOI] [PubMed] [Google Scholar]
  22. Himmelfarb H. J., Maicas E., Friesen J. D. 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. doi: 10.1128/mcb.5.4.816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hughes V., Müller A., Stark M. J., Cohen P. T. Both isoforms of protein phosphatase Z are essential for the maintenance of cell size and integrity in Saccharomyces cerevisiae in response to osmotic stress. Eur J Biochem. 1993 Aug 15;216(1):269–279. doi: 10.1111/j.1432-1033.1993.tb18142.x. [DOI] [PubMed] [Google Scholar]
  24. Kim Y., Huang J., Cohen P., Matthews H. R. Protein phosphatases 1, 2A, and 2C are protein histidine phosphatases. J Biol Chem. 1993 Sep 5;268(25):18513–18518. [PubMed] [Google Scholar]
  25. Kushnirov V. V., Ter-Avanesyan M. D., Telckov M. V., Surguchov A. P., Smirnov V. N., Inge-Vechtomov S. G. Nucleotide sequence of the SUP2 (SUP35) gene of Saccharomyces cerevisiae. Gene. 1988 Jun 15;66(1):45–54. doi: 10.1016/0378-1119(88)90223-5. [DOI] [PubMed] [Google Scholar]
  26. Lee K. S., Hines L. K., Levin D. E. A pair of functionally redundant yeast genes (PPZ1 and PPZ2) encoding type 1-related protein phosphatases function within the PKC1-mediated pathway. Mol Cell Biol. 1993 Sep;13(9):5843–5853. doi: 10.1128/mcb.13.9.5843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Liu H., Krizek J., Bretscher A. Construction of a GAL1-regulated yeast cDNA expression library and its application to the identification of genes whose overexpression causes lethality in yeast. Genetics. 1992 Nov;132(3):665–673. doi: 10.1093/genetics/132.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mann D. J., Dombrádi V., Cohen P. T. Drosophila protein phosphatase V functionally complements a SIT4 mutant in Saccharomyces cerevisiae and its amino-terminal region can confer this complementation to a heterologous phosphatase catalytic domain. EMBO J. 1993 Dec;12(12):4833–4842. doi: 10.1002/j.1460-2075.1993.tb06173.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ohkura H., Kinoshita N., Miyatani S., Toda T., Yanagida M. The fission yeast dis2+ gene required for chromosome disjoining encodes one of two putative type 1 protein phosphatases. Cell. 1989 Jun 16;57(6):997–1007. doi: 10.1016/0092-8674(89)90338-3. [DOI] [PubMed] [Google Scholar]
  30. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Genetic applications of yeast transformation with linear and gapped plasmids. Methods Enzymol. 1983;101:228–245. doi: 10.1016/0076-6879(83)01017-4. [DOI] [PubMed] [Google Scholar]
  31. Ozawa K., Murakami Y., Eki T., Yokoyama K., Soeda E., Hoshino S., Ui M., Hanaoka F. Mapping of the human GSPT1 gene, a human homolog of the yeast GST1 gene, to chromosomal band 16p13.1. Somat Cell Mol Genet. 1992 Mar;18(2):189–194. doi: 10.1007/BF01233164. [DOI] [PubMed] [Google Scholar]
  32. Posas F., Casamayor A., Ariño J. The PPZ protein phosphatases are involved in the maintenance of osmotic stability of yeast cells. FEBS Lett. 1993 Mar 8;318(3):282–286. doi: 10.1016/0014-5793(93)80529-4. [DOI] [PubMed] [Google Scholar]
  33. Posas F., Casamayor A., Morral N., Ariño J. Molecular cloning and analysis of a yeast protein phosphatase with an unusual amino-terminal region. J Biol Chem. 1992 Jun 15;267(17):11734–11740. [PubMed] [Google Scholar]
  34. Posas F., Clotet J., Muns M. T., Corominas J., Casamayor A., Ariño J. The gene PPG encodes a novel yeast protein phosphatase involved in glycogen accumulation. J Biol Chem. 1993 Jan 15;268(2):1349–1354. [PubMed] [Google Scholar]
  35. Reed K. C., Mann D. A. Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 1985 Oct 25;13(20):7207–7221. doi: 10.1093/nar/13.20.7207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ronne H., Carlberg M., Hu G. Z., Nehlin J. O. Protein phosphatase 2A in Saccharomyces cerevisiae: effects on cell growth and bud morphogenesis. Mol Cell Biol. 1991 Oct;11(10):4876–4884. doi: 10.1128/mcb.11.10.4876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Roy A., Lu C. F., Marykwas D. L., Lipke P. N., Kurjan J. The AGA1 product is involved in cell surface attachment of the Saccharomyces cerevisiae cell adhesion glycoprotein a-agglutinin. Mol Cell Biol. 1991 Aug;11(8):4196–4206. doi: 10.1128/mcb.11.8.4196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Russo P., Li W. Z., Hampsey D. M., Zaret K. S., Sherman F. Distinct cis-acting signals enhance 3' endpoint formation of CYC1 mRNA in the yeast Saccharomyces cerevisiae. EMBO J. 1991 Mar;10(3):563–571. doi: 10.1002/j.1460-2075.1991.tb07983.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sharp P. M., Tuohy T. M., Mosurski K. R. Codon usage in yeast: cluster analysis clearly differentiates highly and lowly expressed genes. Nucleic Acids Res. 1986 Jul 11;14(13):5125–5143. doi: 10.1093/nar/14.13.5125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Song J. M., Liebman S. W. Allosuppressors that enhance the efficiency of omnipotent suppressors in Saccharomyces cerevisiae. Genetics. 1987 Mar;115(3):451–460. doi: 10.1093/genetics/115.3.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  42. Stansfield I., Grant G. M., Akhmaloka, Tuite M. F. 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. doi: 10.1111/j.1365-2958.1992.tb01782.x. [DOI] [PubMed] [Google Scholar]
  43. Stinchcomb D. T., Mann C., Davis R. W. Centromeric DNA from Saccharomyces cerevisiae. J Mol Biol. 1982 Jun 25;158(2):157–190. doi: 10.1016/0022-2836(82)90427-2. [DOI] [PubMed] [Google Scholar]
  44. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035–1039. doi: 10.1073/pnas.76.3.1035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sutton A., Immanuel D., Arndt K. T. The SIT4 protein phosphatase functions in late G1 for progression into S phase. Mol Cell Biol. 1991 Apr;11(4):2133–2148. doi: 10.1128/mcb.11.4.2133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tassan J. P., 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. doi: 10.1128/mcb.13.5.2815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Vincent A., Petes T. D. Mitotic and meiotic gene conversion of Ty elements and other insertions in Saccharomyces cerevisiae. Genetics. 1989 Aug;122(4):759–772. doi: 10.1093/genetics/122.4.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wek R. C., Cannon J. F., Dever T. E., Hinnebusch A. G. Truncated protein phosphatase GLC7 restores translational activation of GCN4 expression in yeast mutants defective for the eIF-2 alpha kinase GCN2. Mol Cell Biol. 1992 Dec;12(12):5700–5710. doi: 10.1128/mcb.12.12.5700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wilson P. G., Culbertson M. R. 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. doi: 10.1016/0022-2836(88)90301-4. [DOI] [PubMed] [Google Scholar]
  50. 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 Genetics are provided here courtesy of Oxford University Press

RESOURCES