13C NMR studies of metabolic pathways regulated by HSP104 in Saccharomyces cerevisiae

HSP104 protein in Saccharomyces cerevisiae is known to provide thermotolerance when induced by various kinds of stresses, such asa mild heat shock, ethanol, and hypoxia. It helps cells survive at an otherwise lethal temperature. Mechanisms by which HSP104 protein works are yet to be elucidated. In order to understand a molecular basis of thermotolerance due to KSP104 protein induced by a mild heat shock, studies on respiratory pathways were carried out in the wild type as well as in the hsp104 deleted mutant. Especially the degree of C-13-acetate incorporation into glutamate-C4 was examined for both strains using C-13-C-13 homonuclear spin coupling measurements, since glutamate is in a rapid equilibrium with alpha-ketoglutarate in the TCA cycle. In addition, the temperature effects on the rate of C-13 incorporation are compared with or without HSP104 protein expressed. Finally, the inhibitory effect of HSP104 on the respiration pathway was confirmed by the measurements of oxygen consumption rates for both strains.