Molecular structure of Saccharomyces cerevisiae Gal1p, a bifunctional galatokinase and transcriptional inducer

James B. Thoden, Christopher A. Sellick, David J. Timson, Richard J. Reece, Hazel M. Holden

Research output: Contribution to journalArticlepeer-review

Abstract

Gal1p of Saccharomyces cerevisiae is capable of performing two independent cellular functions. First, it is a key enzyme in the Leloir pathway for galactose metabolism where it catalyzes the conversion of α-D-galactose to galactose 1-phosphate. Second, it has the capacity to induce the transcription of the yeast GAL genes in response to the organism being challenged with galactose as the sole source of carbon. This latter function is normally performed by a highly related protein, Gal3p, but in its absence Gal1p can induce transcription, albeit inefficiently, both in vivo and in vitro. Here we report the x-ray structure of Gal1p in complex with α-D-galactose and Mg-adenosine 5′-(β,γ-imido)triphosphate (AMPPNP) determined to 2.4 Å resolution. Overall, the enzyme displays a marked bilobal appearance with the active site being wedged between dis-tinct N- and C-terminal domains. Despite being considerably larger than other galactokinases, Gal1p shares a similar molecular architecture with these enzymes as well as with other members of the GHMP superfamily. The extraordinary levels of similarity between Gal1p and Gal3p (∼70% amino acid identity and ∼90% similarity) have allowed a model for Gal3p to be constructed. By identifying the locations of mutations of Gal3p that result in altered transcriptional properties, we suggest potential models for Gal3p function and mechanisms for its interaction with the transcriptional inhibitor Gal80p. The GAL genetic switch has long been regarded as a paradigm for the control of gene expression in eukaryotes. Understanding the manner in which two of the proteins that function in transcriptional regulation interact with one another is an important step in determining the overall molecular mechanism of this switch.

Original languageEnglish
Pages (from-to)36905-36911
Number of pages7
JournalJournal of Biological Chemistry
Volume280
Issue number44
DOIs
Publication statusPublished - 4 Nov 2005

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