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In the yeast Saccharomyces cerevisiae, glucose induces expression of the hexose transporter (HXT) genes by inhibiting the repressor function of the transcription factor Rgt1. We have previously shown that Rgt1 binds to the HXT gene promoters only in the absence of glucose. In the presence of glucose, Rgt1 becomes phosphorylated and is unable to bind to the HXT promoters and repress their transcription. We report that Rgt1 interacts with Std1 and Mth1 in a yeast two-hybrid assay and co-immunoprecipitates with both proteins in vivo only when glucose is absent. In addition, we demonstrate that repression of HXT gene expression by Rgt1 is abolished in the std1 mth1 double mutant. While Rgt1 is normally phosphorylated only in the presence of high concentrations of glucose, it is constitutively modified in the std1 mth1 double mutant. Based on these data, we conclude that, in the absence of glucose …