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J Biol Chem, Vol. 274, Issue 39, 28026-28034, September 24, 1999

Overlapping Positive and Negative GATA Factor Binding Sites Mediate Inducible DAL7 Gene Expression in Saccharomyces cerevisiae

From the Department of Microbiology and Immunology, University of Tennessee, Memphis, Tennessee 38163

Allantoin pathway gene expression in Saccharomyces cerevisiae responds to two different environmental stimuli. The expression of these genes is induced in the presence of allantoin or its degradative metabolites and repressed when a good nitrogen source (e.g. asparagine or glutamine) is provided. Three types of cis-acting sites and trans-acting factors are required for allantoin pathway gene transcription as follows: (i) UAS_NTR element associated with the transcriptional activators Gln3p and Gat1p, (ii) URS_GATA element associated with the repressor Dal80p, and (iii) UIS_ALL element associated with the Dal82 and Dal81 proteins required for inducer-dependent transcription. Most of the work leading to the above conclusions has employed inducer-independent allantoin pathway genes (e.g. DAL5 and DAL3). The purpose of this work is to extend our understanding of these elements and their roles to inducible allantoin pathway genes using the DAL7 (encoding malate synthase) as a model. We show that eight distinct cis-acting sites participate in the process as follows: a newly identified GC-rich element, two UAS_NTR, two UIS_ALL, and three URS_GATA elements. The two GATA-containing UAS_NTR elements are coincident with two of the three GATA sequences that make up the URS_GATA elements. The remaining URS_GATA GATA sequence, however, is not a UAS_NTR element but appears to function only in repression. The data provide insights into how these cis- and trans-acting factors function together to accomplish the regulated expression of the DAL7 gene that is observed in vivo.

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