The SNF1 kinase complex from Saccharomyces cerevisiae phosphorylates the transcriptional repressor protein Mig1p in vitro at four sites within or near regulatory domain 1

FEBS Letters

Hardie, D.Grahame¹  Wilson, Wayne A.¹  Davies, Stephen P.¹  Carling, David²  Smith, Fiona C.² 

[1] Biochemistry Department, Dundee University, MSI/WTB Complex, Dow Street, Dundee DD1 5EH, UK;Cellular Stress Group, MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK

关键词: SNF1;    Mig1p;    Glucose repression;    Protein phosphorylation;    AMPK;    AMP-activated protein kinase;    GST;    glutathione-S-transferase;    PCR;    polymerase chain reaction;    TCA;    trichloroacetic acid;    TFA;    trifluoroacetic acid;    SDS;    sodium dodecyl sulfate;    VP16;    viral protein 16;   

DOI  :  10.1016/S0014-5793(99)00725-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Mig1p is a zinc finger protein required for repression of glucose-regulated genes in budding yeast. On removal of medium glucose, gene repression is relieved via a mechanism that requires the SNF1 protein kinase complex. We show that Mig1p expressed as a glutathione-S-transferase fusion in bacteria is readily phosphorylated by the SNF1 kinase in vitro. Four phosphorylation sites were identified, i.e. Ser-222, Ser-278, Ser-311 and Ser-381. The latter three are exact matches to the recognition motif we previously defined for SNF1 and lie within regions shown to be required for SNF1-dependent derepression and nuclear-to-cytoplasmic translocation.

【 授权许可】

Unknown