Isolation Methods for High-Pressure Growth Mutant in Yeast

  • Fumiyoshi Abe
Reference work entry

Isolation of High-Pressure Growth Mutants from Tryptophan-Auxotrophic Strains of the Yeast Saccharomyces cerevisiae

As mentioned in Chap. 5.4, wild-type strains of S. cerevisiae with tryptophan auxotrophy (Trp) are unable to grow under high pressure (e.g., 15–25 MPa, 25°C) and low temperature (e.g., 0.1 MPa, 10–15°C). This is because high pressure and low temperature compromise tryptophan uptake by inhibiting the permease activity and promoting ubiquitin-dependent degradation of the permeases (Abe 2007). Mutants capable of growth under high pressure, designated HPG (high-pressure growth) mutants (referred to as the HPG phenotype, hereafter), are isolated from tryptophan-auxotrophic strains (Abe and Iida 2003). The characterization of the HPGmutants and identification of relevant genes allow us to unravel the complex regulatory mechanism of the yeast tryptophan permeases Tat1 and Tat2 with respect to ubiquitination, deubiquitination, and endocytic trafficking in the cell. In this...


High Hydrostatic Pressure Amino Acid Uptake Tryptophan Uptake Dual Excitation Wavelength Tryptophan Permease 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. Abe F (2007) Exploration of the effects of high hydrostatic pressure on microbial growth, physiology and survival: perspectives from piezophysiology. Biosci Biotechnol Biochem 71:2347–2357PubMedCrossRefGoogle Scholar
  2. Abe F, Horikoshi K (1995) Hydrostatic pressure promotes the acidification of vacuoles in Saccharomyces cerevisiae. FEMS Microbiol Lett 130:307–312PubMedCrossRefGoogle Scholar
  3. Abe F, Horikoshi K (1997) Vacuolar acidification in Saccharomyces cerevisiae induced by elevated hydrostatic pressure is transient and is mediated by vacuolar H+-ATPase. Extremophiles 1:89–93PubMedCrossRefGoogle Scholar
  4. Abe F, Horikoshi K (1998) Analysis of intracellular pH in the yeast Saccharomyces cerevisiae under elevated hydrostatic pressure: a study in baro- (piezo-) physiology. Extremophiles 2:223–228PubMedCrossRefGoogle Scholar
  5. Abe F, Iida H (2003) Pressure-induced differential regulation of the two tryptophan permeases Tat1 and Tat2 by ubiquitin ligase Rsp5 and its binding proteins, Bul1 and Bul2. Mol Cell Biol 23:7566–7584PubMedCrossRefGoogle Scholar
  6. Abe F, Minegishi H (2008) Global screening of genes essential for growth in high-pressure and cold environments: searching for basic adaptive strategies using a yeast deletion library. Genetics 178:851–872PubMedCrossRefGoogle Scholar
  7. Giaever G, Chu AM, Ni L, Connelly C, Riles L, Veronneau S, Dow S, Lucau-Danila A, Anderson K, Andre B, Arkin AP, Astromoff A, El-Bakkoury M, Bangham R, Benito R, Brachat S, Campanaro S, Curtiss M, Davis K, Deutschbauer A, Entian KD, Flaherty P, Foury F, Garfinkel DJ, Gerstein M, Gotte D, Guldener U, Hegemann JH, Hempel S, Herman Z, Jaramillo DF, Kelly DE, Kelly SL, Kotter P, LaBonte D, Lamb DC, Lan N, Liang H, Liao H, Liu L, Luo C, Lussier M, Mao R, Menard P, Ooi SL, Revuelta JL, Roberts CJ, Rose M, Ross-Macdonald P, Scherens B, Schimmack G, Shafer B, Shoemaker DD, Sookhai-Mahadeo S, Storms RK, Strathern JN, Valle G, Voet M, Volckaert G, Wang CY, Ward TR, Wilhelmy J, Winzeler EA, Yang Y, Yen G, Youngman E, Yu K, Bussey H, Boeke JD, Snyder M, Philippsen P, Davis RW, Johnston M (2002) Functional profiling of the Saccharomyces cerevisiae genome. Nature 418:387–391PubMedCrossRefGoogle Scholar
  8. Haworth RS, Lemire BD, Crandall D, Cragoe EJ Jr, Fliegel L (1991) Characterisation of proton fluxes across the cytoplasmic membrane of the yeast Saccharomyces cerevisiae. Biochim Biophys Acta 1098:79–89PubMedCrossRefGoogle Scholar
  9. Kitamura Y, Itoh T (1987) Reaction volume of protonic ionization for buffering agents. Prediction of pressure dependence of pH and pOH. J Solution Chem 16:715–725CrossRefGoogle Scholar
  10. Nagayama A, Kato C, Abe F (2004) The N- and C-terminal mutations in tryptophan permease Tat2 confer cell growth in Saccharomyces cerevisiae under high-pressure and low-temperature conditions. Extremophiles 8:143–149PubMedCrossRefGoogle Scholar
  11. Nelissen B, De Wachter R, Goffeau A (1997) Classification of all putative permeases and other membrane plurispanners of the major facilitator superfamily encoded by the complete genome of Saccharomyces cerevisiae. FEMS Microbiol Rev 21:113–134PubMedCrossRefGoogle Scholar
  12. Preston RA, Murphy RF, Jones EW (1989) Assay of vacuolar pH in yeast and identification of acidification-defective mutants. Proc Natl Acad Sci USA 86:7027–7031PubMedCrossRefGoogle Scholar
  13. Schmidt A, Hall MN, Koller A (1994) Two FK506 resistance-conferring genes in Saccharomyces cerevisiae, TAT1 and TAT2, encode amino acid permeases mediating tyrosine and tryptophan uptake. Mol Cell Biol 14:6597–6606PubMedGoogle Scholar

Copyright information

© Springer 2011

Authors and Affiliations

  1. 1.Molecular Evolution and Adaptation Research, Institute of BiogeosciencesJapan Agency for Marine-Earth Science and Technology (JAMSTEC)YokosukaJapan
  2. 2.Aoyama Gakuin UniversitySagamiharaJapan

Personalised recommendations