Regulation of THI4 (MOL1), a thiamine-biosynthetic gene of Saccharomyces cerevisiae

Yeast
U M PraekeltP Meacock

Abstract

THI4, a Saccharomyces cerevisiae gene originally identified as a result of transient expression in molasses medium and named MOL1 is regulated by thiamine. Using a THI4 promoter-lacZ fusion on a centromeric yeast vector, we have shown that the THI4 is completely repressed throughout batch culture by thiamine at a concentration around 1 microM, but shows high level constitutive expression in thiamine-free medium. The transient expression pattern observed in molasses medium can be mimicked by the addition of 0.15 microM-thiamine to defined minimal medium. Cells grown in thiamine-free medium have an intracellular thiamine concentration of around 9 pmol/10(7) cells. A low level (1 microM) of exogenous thiamine is completely sequestered from the medium within 30 min; intracellular thiamine concentrations rise rapidly, followed by a gradual decrease as a result of dilution during growth. A saturating extracellular level of thiamine leads to a maximal intracellular concentration of around 1600 pmol/10(7) cells, at which point the transport system is shut down. After transfer from repressing to non-repressing medium, THI4 becomes induced when the intracellular concentration of thiamine falls to 20 pmol/10(7) cells. A thi4::URA3 disrupt...Continue Reading

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Related Concepts

THI4 protein, S cerevisiae
Feedback - System Communication
Yeast Proteins
Genes, Fungal
Molasses
Chimeric Proteins, Recombinant
Saccharomyces cerevisiae
Thiamine Mononitrate
Gene Expression Regulation, Fungal
Saccharomyces cerevisiae Proteins

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