Skip to main content
Log in

HSP12, a new small heat shock gene of Saccharomyces cerevisiae: Analysis of structure, regulation and function

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

We have isolated a new small heat shock gene, HSP12, from Saccharomyces cerevisiae. It encodes a polypeptide of predicted Mr 12 kDa, with structural similarity to other small heat shock proteins. HSP12 gene expression is induced several hundred-fold by heat shock and on entry into stationary phase. HSP12 mRNA is undetectable during exponential growth in rich medium, but low levels are present when cells are grown in minimal medium. Analysis of HSP12 expression in mutants affected in cAMP-dependent protein phosphorylation suggests that the gene is regulated by cAMP as well as heat shock. A disruption of the HSP12 coding region results in the loss of an abundant 14.4 kDa protein present in heat shocked and stationary phase cells. It also leads to the induction of the heat shock response under conditions normally associated with low-level HSP12 expression. The HSP12 disruption has no observable effect on growth at various temperatures, nor on the ability to acquire thermotolerance.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ananthan J, Goldberg AL, Voellmy R (1986) Abnormal proteins serve as eukaryotic stress signals and trigger the activation of heat shock genes. Science 232:522–524

    Google Scholar 

  • Bennetzen JL, Hall BD (1982) Codon selection in yeast. J Biol Chem 257:3029–3031

    Google Scholar 

  • Berger EM, Woodward MP (1983) Small heat shock proteins in Drosophila may confer thermal tolerance. Exp Cell Res 147:437–442

    Google Scholar 

  • Borkovich KA, Farrelly FW, Finkelstein DB, Taulien J, Lindquist S (1989) Hsp82 is an essential protein that is required in higher concentrations for growth of cells at higher temperatures. Mol Cell Biol 9:3919–3930

    Google Scholar 

  • Bossier P, Fitch IA, Boucherie H, Tuite MF (1989) Structure and expression of a yeast gene encoding the small heat shock protein, Hsp26. Gene 78:323–330

    Google Scholar 

  • Botstein D, Falco SC, Stewart SE, Brennan M, Scherer S, Stinchcomb DT, Struhl K, Davies RW (1979) Sterile host yeasts (SHY): A eukaryotic system of biological containment for recombinant DNA experiments. Gene 8:17–24

    Google Scholar 

  • Chirico WJ, Waters MG, Blobel G (1988) 70K heat shock related proteins stimulate protein translocation into microsomes. Nature 332:805–810

    Google Scholar 

  • Cigan AM, Donahue TF (1987) Sequence and structural features associated with translational initiator regions in yeast — a review. Gene 59:1–18

    Google Scholar 

  • Craig EA, Jacobsen K (1984) Mutations of the heat inducible 70 kilodalton genes of yeast confer temperature sensitive growth. Cell 38:841–849

    Google Scholar 

  • Craig EA, Jacobsen K (1985) Mutations in cognate genes of Saccharomyces cerevisiae hsp70 result in reduced growth rates at low temperatures. Mol Cell Biol 5:3517–3524

    Google Scholar 

  • Deshaies RJ, Koch BD, Werner-Washburne M, Craig EA, Schekman R (1988) A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides. Nature 332:800–805

    Google Scholar 

  • DiDomenico BJ, Bugaisky GE, Lindquist S (1982) The heat shock response is self-regulated at both the transcriptional and post-transcriptional levels. Cell 31:593–603

    Google Scholar 

  • Dobson MJ, Tuite MF, Roberts NA, Kingsman AJ, Kingsman SM (1982) Conservation of high efficiency promoter sequences in Saccharomyces cerevisiae. Nucleic Acids Res 10:2625–2637

    Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13

    Google Scholar 

  • Finley D, Oezkaynak E, Varshavsky A (1987) The yeast polyubiquitin gene is essential for resistance to high temperatures, starvation, and other stresses. Cell 48:1035–1046

    Google Scholar 

  • Frischauf AM, Lehrach H, Poustka A, Murray NM (1983) Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170:827–842

    Google Scholar 

  • Fujita T, Ohno S, Yasumitsu H, Taniguchi T (1985) Delimitation and properties of DNA sequences required for the regulated expression of human interferon-β gene. Cell 41:489–496

    Google Scholar 

  • Grant CM, Firoozan M, Tuite MF (1989) Mistranslation induces the heat-shock response in the yeast Saccharomyces cerevisiae. Mol Microbiol 3:215–220

    Google Scholar 

  • Hames BD (1981) In: Hames BD, Rickwood D (eds) Gel electrophoresis of proteins. A practical approach. IRL Press, Oxford, pp 1–91

    Google Scholar 

  • Iida H, Yahara I (1984) Durable synthesis of high molecular weight heat shock proteins in Go cells of the yeast and other eukaryotes. J Cell Biol 99:199–209

    Google Scholar 

  • Ingolia TD, Craig EA (1982) Four small Drosophila heat shock proteins are related to each other and to mammalian α-crystallin. Proc Natl Acad Sci USA 79:2360–2364

    Google Scholar 

  • Kyte J, Doolittle RF (1982) A simple method for displaying the hydrophobic character of a protein. J Mol Biol 157:105–132

    Google Scholar 

  • Lindquist S (1986) The heat-shock response. Annu Rev Biochem 55:1151–1191

    Google Scholar 

  • Lindquist S, Craig EA (1988) The heat-shock proteins. Annu Rev Genet 22:631–677

    Google Scholar 

  • Loomis WF, Wheeler SA (1982) Chromatin-associated heat shock proteins of Dictyostelium. Dev Biol 90:412–418

    Google Scholar 

  • Makara MP, Henson JM (1985) BLOTTO for dried agarose gels. Focus 8:14–15

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Matsumoto K, Uno I, Ishikawa T (1985) Genetic analysis of the role of cAMP in yeast. Yeast 1:15–24

    Google Scholar 

  • McAlister L, Finkelstein DB (1980) Heat shock proteins and thermal resistance in yeast. Biochem Biophys Res Commun 93:819–824

    Google Scholar 

  • Normington K, Kohno K, Kozutsumi Y, Gething M-J, Sambrook J (1989) S. cerevisiae encodes an essential protein homologous in sequence and function to mammalian Pip. Cell 57:1233–1236

    Google Scholar 

  • Oezkaynak E, Finley D, Solomon MJ, Varshavsky A (1987) The yeast ubiquitin genes: a family of natural gene fusions. EMBO J 6:1429–1439

    Google Scholar 

  • Ostermann J, Horwich AL, Neupert W, Hard F-U (1989) Protein folding in mitochondria requires complex formation with hsp60 and ATP hydrolysis. Nature 341:125–130

    Google Scholar 

  • Pelham HRB (1989) Heat shock and the sorting of luminal ER proteins. EMBO J 8:3171–3176

    Google Scholar 

  • Petko L, Lindquist S (1986) Hsp26 is not required for growth at high temperatures, nor for thermotolerance, spore development, or germination. Cell 45:885–894

    Google Scholar 

  • Proudfoot NJ, Brownlee GG (1976) 3′Non-coding region sequences in eukaryotic messenger RNA. Nature 263:211–214

    Google Scholar 

  • Raschke E, Baumann G, Schoeffl F (1988) Nucleotide sequence analysis of soybean small heat shock protein genes belonging to two different multigene families. J Mol Biol 199:549–557

    Google Scholar 

  • Riddihough G, Pelham HRB (1986) Activation of the Drosophila hsp27 promoter by heat shock and by ecdysone involves independent and remote regulatory sequences. EMBO J 5:1653–1658

    Google Scholar 

  • Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–210

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sarokin L, Carlson M (1986) Short repeated elements in the upstream regulatory region of the SUC2 gene of Saccharomyces cerevisiae. Mol Cell Biol 6:2324–2333

    Google Scholar 

  • Schlesinger MJ (1986) Heat shock proteins: The search for functions. J Cell Biol 103:321–325

    Google Scholar 

  • Sharp PM, Tuohy TMF, Mosurski KR (1986) Codon usage in yeast: cluster analysis clearly differentiates highly and lowly expressed genes. Nucleic Acids Res 14:5125–5143

    Google Scholar 

  • Sherman F, Fink GR, Hicks JB (1986) Laboratory course manual for methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sorger PK, Pelham HRB (1987) Purification and characterisation of a heat shock element binding protein from yeast. EMBO J 6:3035–3041

    Google Scholar 

  • Sorger PK, Pelham HRB (1988) Yeast heat shock factor is an essential DNA-binding protein that exhibits temperatur dependent phosphorylation. Cell 54:855–864

    Google Scholar 

  • Struhl K (1986) In: Reznikoff W, Gold L (eds) From gene to protein: Steps dictating the maximal level of gene expression. Butterworth Publishing Co, Boston, pp 35–78

    Google Scholar 

  • Struhl K (1987) Promoters, activator proteins, and the mechanism of transcriptional initiation in yeast. Cell 49:295–297

    Google Scholar 

  • Tanaka K, Matsumoto K, Toh-e A (1988) Dual regulation of the expression of the polyubiquitin gene by cyclic AMP and heat shock in yeast. EMBO J 7:495–502

    Google Scholar 

  • Tuite MF, Bossier P, Fitch IT (1988) A highly conserved sequence in yeast heat shock gene promoters. Nucleic Acids Res 16:11845

    Google Scholar 

  • Vieira J, Messing J (1982) The pUC plasmids, and M13mp7-derived system for insertion mutagenesis and sequencing with universal primers. Gene 19:259–268

    Google Scholar 

  • 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

    Google Scholar 

  • Werner-Washburne M, Becker J, Kosic-Smithers J, Craig EA (1989) Yeast Hsp70 RNA levels vary in response to the physiological status of the cell. J Bacteriol 171:2680–2688

    Google Scholar 

  • Wu BJ, Kingston RE, Morimoto RI (1986) Human HSP70 promoter contains at least two distinct regulatory domains. Proc Natl Acad Sci USA 83:629–633

    Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119

    Google Scholar 

  • Zaret KS, Sherman F (1982) DNA sequence required for efficient transcription termination in yeast. Cell 28:563–573

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by C.P. Hollenberg

Rights and permissions

Reprints and permissions

About this article

Cite this article

Praekelt, U.M., Meacock, P.A. HSP12, a new small heat shock gene of Saccharomyces cerevisiae: Analysis of structure, regulation and function. Mol Gen Genet 223, 97–106 (1990). https://doi.org/10.1007/BF00315801

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00315801

Key words

Navigation