Thiamin metabolism and thiamin diphosphate-dependent enzymes in the yeast Saccharomyces cerevisiae: genetic regulation

Abstract

The yeast Saccharomyces cerevisiae utilises external thiamin for the production of thiamin diphosphate (ThDP) or can synthesise the cofactor itself. Prior to uptake into the cell thiamin phosphates are first hydrolysed and thiamin is taken up as free vitamin which is then pyrophosphorylated by a pyrophosphokinase. Synthesis of ThDP starts with the production of hydroxyethylthiazole and hydroxymethylpyrimidine. Those are linked to yield thiamin phosphate which is hydrolysed to thiamin and subsequently pyrophosphorylated. The THI genes encoding the enzymes of these final steps of ThDP production and of thiamin utilisation have been identified. Their expression is controlled by the level of thiamin and a number of regulatory proteins involved in regulated expression of the THI genes are known. However, the molecular details of the regulatory circuits need to be deciphered. Since the nucleotide sequence of the entire yeast genome is known we can predict the number of ThDP-dependent enzymes in S. cerevisiae. Eleven such proteins have been found: pyruvate decarboxylase (Pdc, three isoforms), acetolactate synthase, a putative alpha-ketoisocaproate decarboxylase with a regulatory role in ThDP synthesis and two proteins of unknown function form the group of Pdc related enzymes. In addition there are two isoforms for transketolase as well as the E1 subunits of pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase. Expression of most of these genes is either induced or repressed by glucose. Surprisingly, it has been found recently that expression of one of the genes for Pdc is repressed by thiamin. In addition, the regulatory protein Pdc2p was shown to be required for high level expression of both the THI and the PDC genes. Apparently, the production of ThDP and of the enzymes using this cofactor is coordinately regulated. Future research will focus on the elucidation of the molecular mechanisms of this novel type of regulation.