Interaction of the repressors Nrg1 and Nrg2 with the Snf1 protein kinase in Saccharomyces cerevisiae.
The Snf1 protein kinase is essential for the transcription of glucose-repressed genes in Saccharomyces cerevisiae. We identified Nrg2 as a protein that interacts with Snf1 in the two-hybrid system. Nrg2 is a C(2)H(2) zinc-finger protein that is homologous to Nrg1, a repressor of the glucose- and Snf1-regulated STA1 (glucoamylase) gene. Snf1 also interacts with Nrg1 in the two-hybrid system and co-immunoprecipitates with both Nrg1 and Nrg2 from cell extracts. A LexA fusion to Nrg2 represses transcription from a promoter containing LexA binding sites, indicating that Nrg2 also functions as a repressor. An Nrg1 fusion to green fluorescent protein is localized to the nucleus, and this localization is not regulated by carbon source. Finally, we show that VP16 fusions to Nrg1 and Nrg2 allow low-level expression of SUC2 in glucose-grown cells, and we present evidence that Nrg1 and Nrg2 contribute to glucose repression of the DOG2 gene. These results suggest that Nrg1 and Nrg2 are direct or indirect targets of the Snf1 kinase and function in glucose repression of a subset of Snf1-regulated genes.
Vyas
V K
VK
Integrated Program in Cellular Biology, Molecular Biology and Biophysical Studies, Columbia University, New York, NY 10032, USA.
Kuchin
S
S
Carlson
M
M
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001665:Binding Sites; D002244:Carbon / Q000378:metabolism; D020134:Catalytic Domain; D002467:Cell Nucleus / Q000378:metabolism; D017931:DNA Primers / Q000378:metabolism; D004268:DNA-Binding Proteins; D004789:Enzyme Activation; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000378:metabolism; D000099160:LexA Repressor Protein
D001426:Bacterial Proteins; D002244:Carbon; D017931:DNA Primers; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D005087:Glucan 1,4-alpha-Glucosidase; D005947:Glucose; D010744:Phosphoric Monoester Hydrolases; D017346:Protein Serine-Threonine Kinases; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D012697:Serine Endopeptidases; D001616:beta-Galactosidase; D000099160:LexA Repressor Protein; C117881:NRG1 protein, S cerevisiae; C435344:Nrg2 protein, S cerevisiae; C071570:SNF1-related protein kinases; C093600:DOG2 protein, S cerevisiae