Understanding the mechanism of glucose-induced relief of Rgt1-mediated repression in yeast.
The yeast Rgt1 repressor inhibits transcription of the glucose transporter (HXT) genes in the absence of glucose. It does so by recruiting the general corepressor complex Ssn6-Tup1 and the HXT corepressor Mth1. In the presence of glucose, Rgt1 is phosphorylated by the cAMP-activated protein kinase A (PKA) and dissociates from the HXT promoters, resulting in expression of HXT genes. In this study, using Rgt1 chimeras that bind DNA constitutively, we investigate how glucose regulates Rgt1 function. Our results show that the DNA-bound Rgt1 constructs repress expression of the HXT1 gene in conjunction with Ssn6-Tup1 and Mth1, and that this repression is lifted when they dissociate from Ssn6-Tup1 in high glucose conditions. Mth1 mediates the interaction between the Rgt1 constructs and Ssn6-Tup1, and glucose-induced downregulation of Mth1 enables PKA to phosphorylate the Rgt1 constructs. This phosphorylation induces dissociation of Ssn6-Tup1 from the DNA-bound Rgt1 constructs, resulting in derepression of HXT gene expression. Therefore, Rgt1 removal from DNA occurs in response to glucose but is not necessary for glucose induction of HXT gene expression, suggesting that glucose regulates Rgt1 function by primarily modulating the Rgt1 interaction with Ssn6-Tup1.
Department of Biochemistry and Molecular Medicine, The George Washington University Medical Center, 2300 Eye Street, Washington, DC 20037, USA.
Adhiraj
A
Roy
Department of Biological Sciences, The University of Southern Mississippi, 118 College Dr., Hattiesburg, MS 39406, USA.
David
D
Jouandot
Department of Microbiology, Southern Illinois University Carbondale, 1125 Lincoln Drive, Carbondale, IL 62901, USA.
Kyu Hong
KH
Cho
Department of Biochemistry and Molecular Medicine, The George Washington University Medical Center, 2300 Eye Street, Washington, DC 20037, USA.
Jeong-Ho
JH
Kim
ChIP, chromatin immunoprecipitation; DNA-binding; ECL, enhanced chemiluminescence; Glucose; HXT; IP, immunoprecipitation; PKA, protein kinase A; Rgt1; Yeast