Isolation Methods for High-Pressure Growth Mutant in Yeast
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...
KeywordsHigh Hydrostatic Pressure Amino Acid Uptake Tryptophan Uptake Dual Excitation Wavelength Tryptophan Permease
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