Regulation of the nucleocytoplasmic distribution of Snf1-Gal83 protein kinase.
Snf1 protein kinase containing the beta subunit Gal83 is localized in the cytoplasm during growth of Saccharomyces cerevisiae cells in abundant glucose and accumulates in the nucleus in response to glucose limitation. Nuclear localization of Snf1-Gal83 requires activation of the Snf1 catalytic subunit and depends on Gal83, but in the snf1Delta mutant, Gal83 exhibits glucose-regulated nuclear accumulation. We show here that the N terminus of Gal83, which is divergent from those of the other beta subunits, is necessary and sufficient for Snf1-independent, glucose-regulated localization. We identify a leucine-rich nuclear export signal in the N terminus and show that export depends on the Crm1 export receptor. We present evidence that catalytically inactive Snf1 promotes the cytoplasmic retention of Gal83 in glucose-grown cells through its interaction with the C terminus of Gal83; cytoplasmic localization of inactive Snf1-Gal83 maintains accessibility to the Snf1-activating kinases. Finally, we characterize the effects of glucose phosphorylation on localization. These studies define roles for Snf1 and Gal83 in determining the nucleocytoplasmic distribution of Snf1-Gal83 protein kinase.
Hedbacker
Kristina
K
Department of Genetics and Development, Columbia University, 701 W. 168th Street, HSC922, New York, NY 10032, USA.
Carlson
Marian
M
D021581:Active Transport, Cell Nucleus; D003847:Deoxyglucose / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005800:Genes, Fungal; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics / Q000378:metabolism; D028884:Karyopherins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D049790:Nuclear Export Signals; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000097606:Exportin 1 Protein
D003847:Deoxyglucose; D004791:Enzyme Inhibitors; D005947:Glucose; D005956:Glucose-6-Phosphate Isomerase; D028884:Karyopherins; D049790:Nuclear Export Signals; D017346:Protein Serine-Threonine Kinases; D018160:Receptors, Cytoplasmic and Nuclear; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D000097606:Exportin 1 Protein; C085569:GAL83 protein, S cerevisiae; C071570:SNF1-related protein kinases