Molecular and General Genetics MGG

, Volume 245, Issue 1, pp 107–116

A new yeast gene, HTR1, required for growth at high temperature, is needed for recovery from mating pheromone-induced G1 arrest

  • Yoshiko Kikuchi
  • Yoshio Oka
  • Mariko Kobayashi
  • Yukifumi Uesono
  • Akio Toh-e
  • Akihiko Kikuchi
Original Paper

Abstract

A new temperature-sensitive mutant of Saccharomyces cerevisiae was isolated. Arrested cells grown at the nonpermissive temperature were of dumb-bell shape and contained large vacuoles. A DNA fragment was cloned based on its ability to complement this temperature sensitivity. The HTR1 gene encodes a putative protein of 93 kDa without significant homology to any known proteins. The gene was mapped between ade5 and lys5 on the left arm of chromosome VII. The phenotype of the gene disruptant appeared to be strain-specific; disruption of the gene in strain W303 caused the cells to become temperature sensitive. The arrested phenotype here was similar to that of the original is mutant and cells in G2/M phase predominated at high temperature. Another disruptant in a strain YPH background grew slowly at high temperature due to slow progression through G2/M phase, and morphologically abnormal (elongated) cells accumulated. A single-copy suppressor that alleviated the temperature-sensitive defects in both strains was identified as MCS1/SSD1. The wild-type strains W303 and YPH are known to carry defective MCS1/SSD1 alleles; hence HTR1 may function redundantly with MCS1/SSD1 to suppress the temperature-sensitive phenotypes. In addition, based on a halo bioassay, the disruptant strains appeared to be defective in recovery from, or adaptive response to G1 arrest mediated by mating pheromone, even at the permissive temperature. Thus the gene has at least two functions and is designated HTR1 (required for high temperature growth and recovery from G1 arrest induced by mating pheromone).

Key words

Saccharomyces cerevisiae ts mutant Recovery HTR1 MCS1/SSD1 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Arndt KT, Styles CA, Fink GR (1989) A suppressor of HIS4 transcriptional defect encodes a protein with homology to the catalytic subunit of protein phosphatases. Cell 56:527–537Google Scholar
  2. Beggs JD (1978) Transformation of yeast by a replicating hybrid plasmid. Nature 275:104–109Google Scholar
  3. Botstein DS, Falco C, Stewart SE, Brennan M, Sherer S, Stinchcomb DT, Struhl K, Davis, RW (1979) Sterile host yeast (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene 8:17–23Google Scholar
  4. Ciejek E, Thorner J (1979) Recovery of S. cerevisiae a cells from G1 arrest by α factor pheromone requires endopeptidase action. Cell 18:623–635Google Scholar
  5. Cole GM, Reed SI (1991) Pheromone-induced phosphorylation of a G protein β subunit in S. cerevisiae is associated with an adaptive response to mating pheromone. Cell 64:703–716Google Scholar
  6. Costigan C, Gehrung S, Snyder M (1992) A synthetic lethal screen identifies SLK1, a novel protein kinase homology implicated in yeast cell morphogenesis and cell growth. Mol Cell Biol 12:1162–1178Google Scholar
  7. Cross FR (1988) DAF1, a mutant gene affecting size control, pheromone arrest, and cell cycle kinetics of Saccharomyces cerevisiae. Mol Cell Biol 8:4675–4684Google Scholar
  8. Cross FR, Tinkelenberg AH (1991) A potential positive feedback loop controlling CLN1 and CLN2 gene expression at the start of the yeast cell cycle. Cell 65:875–883Google Scholar
  9. Dietzel C, Kurjan J (1987) Pheromonal regulation and sequence of the Saccharomyces cerevisiae SST2 gene: a model for desensitization to pheromone. Mol Cell Biol 7:4169–4177Google Scholar
  10. Dirick L, Nasmyth K (1991) positive feedback in the activation of G1 cyclins in yeast. Nature 351:754–757Google Scholar
  11. Healy AM, Zolnierowicz S, Stapleton AE, Goebl M, DePaoli-Roach AA, Pringle JR (1991) CDC55, a Saccharomyces cerevisiae gene involved in cellular morphogenesis: identification, characterization, and homology to the B subunit of mammalian type 2A protein phosphatase. Mol Cell Biol 11:5767–5780Google Scholar
  12. Irie K, Nomoto S, Miyajima I, Matsumoto KJ (1991) SGV1 encodes a CDC28/cdc2-related kinase required for a Gα subunit-mediated adaptive response to pheromone in S. cerevisiae. Cell 65:785–795Google Scholar
  13. Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168Google Scholar
  14. Kikuchi Y, Shimatake H, Kikuchi A (1988) A yeast gene required for the G1-to-S transition encodes a protein containing an A-kinase target site and GTPase domain. EMBO J 7:1175–1182Google Scholar
  15. Kinoshita N, Goebl M, Yanagida M (1991) The fission yeast dis3 + gene encodes a 110-kDa essential protein implicated in mitotic control. Mol Cell Biol 11:5839–5847Google Scholar
  16. Konopka JB, Jenness DD and Hartwell LH (1988) The C-terminus of the Saccharomyces cerevisiae α-pheromone receptor mediates an adaptive response to pheromone. Cell 54:609–618Google Scholar
  17. Kronstad JW, Holly JA, Mackay VL (1987) A yeast operator overlaps an upstream activation site. Cell 50:369–377Google Scholar
  18. Kuo C-L, Campbell JL (1983) Cloning of Saccharomyces cerevisiae DNA replication genes: isolation of the CDC8 gene and two genes that compensate for the cdc8-1 mutation. Mol Cell Biol 3:1730–1737Google Scholar
  19. Lee KS, Irie K, Gotoh Y, Watanabe Y, Araki H, Nishida E, Matsumoto K, Levin DE (1993) A yeast mitogen-activated protein kinase homolog (Mpk1p) mediates signalling by protein kinase C. Mol Cell Biol 13:3067–3075Google Scholar
  20. Marsh L, Neiman AM, Herskowitz I (1991) Signal transduction during pheromone response in yeast. Annu Rev Cell Biol p699–728Google Scholar
  21. Miyajima I, Arai K, Matsumoto K (1989) GPA1 Val-50 mutation in the mating-factor signaling pathway in Saccharomyces cerevisiae. Mol Cell Biol 9:2289–2297Google Scholar
  22. Nash R, Tokiwa G, Anand S, Erickson K, Futcher B (1988) The WHII + gene of Saccharomyces cerevisiae tethers cell division to cell size and is a cyclin homolog. EMBO J 7:4335–4346Google Scholar
  23. Nigg EA (1993) Cellular substrates of p34cdc2 and its companion cyclin-dependent kinases. Trends Cell Biol 3:296–301Google Scholar
  24. Pringle JR, Hartwell LH (1981) The Saccharomyces cerevisiae cell cycle. In: Broach JE, Pringle JR, Jones EW (eds) The molecular biology of the yeast Saccharomyces: Life cycle and inheritance. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 97–142Google Scholar
  25. Pringle JR, Preston RA, Adams AEM, Stearns T, Drubin DG, Haarer BK, Jones EW (1989) Fluorescence microscopy methods for yeast. Methods Enzymol 31:357–435Google Scholar
  26. Reneke JE, Blumer KJ, Courchesne WE, Thorner J (1988) The carboxy-terminal segment of the yeast a-factor receptor is a regulatory domain. Cell 55:221–234Google Scholar
  27. Ronne H, Carlberg M, Hu G-Z, Nehlin JO (1991) Protein phosphatase 2A in Saccharomyces cerevisiae: effects on cell growth and bud morphogenesis. Mol Cell Biol 11:4876–4884Google Scholar
  28. Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211Google Scholar
  29. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  30. Sherman F, Fink GR, Hicks JM (1986) Laboratory course manual for methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  31. Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122:19–27Google Scholar
  32. Surana U, Robitsch H, Price C, Schuster T, Fitch I, Futcher AB, Nasmyth K, (1991) The role of CDC28 and cyclins during mitosis in the budding yeast S. cerevisiae. Cell 65:145–161Google Scholar
  33. Sutton A, Immanuel D, Arndt KT (1991) The SIT4 protein phosphatase functions in late G1 for progression into S phase. Mol Cell Biol 11:2133–2148Google Scholar
  34. Uesono Y, Fujita A, Toh-e A, Kikuchi Y (1994) The MCS1/SSD1/SRK1/SSL1 gene is involved in stable maintenance of the chromosome in yeast. Gene 143:135–138Google Scholar
  35. Wilson RB, Brenner AA, White TB, Engler MJ, Gaughran JP, Tatchell K (1991) The Saccharomyces cerevisiae SRK1 gene, a suppressor of bcy1 and ins1, may be involved in protein phosphatase function. Mol Cell Biol 11:3369–3373Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Yoshiko Kikuchi
    • 1
  • Yoshio Oka
    • 2
  • Mariko Kobayashi
    • 2
  • Yukifumi Uesono
    • 1
  • Akio Toh-e
    • 1
  • Akihiko Kikuchi
    • 2
  1. 1.Department of Plant Sciences, Graduate School of ScienceThe University of TokyoBunkyo-ku, TokyoJapan
  2. 2.Mitsubishi Kasei Institute of Life SciencesMachida-shi, TokyoJapan

Personalised recommendations