Abstract
The essential, rapamycin-sensitive TOR kinases regulate a diverse set of cell growth-related readouts in response to nutrients. Thus, the yeast TOR proteins function as nutrient sensors, in particular as sensors of nitrogen and possibly carbon. However, the nutrient metabolite(s) that acts upstream of TOR is unknown. We investigated the role of glutamine, a preferred nitrogen source and a key intermediate in yeast nitrogen metabolism, as a possible regulator of TOR. We show that the glutamine synthetase inhibitor L-methionine sulfoximine (MSX) specifically provokes glutamine depletion in yeast cells. MSX-induced glutamine starvation caused nuclear localization and activation of the TOR-inhibited transcription factors GLN3, RTG1, and RTG3, all of which mediate glutamine synthesis. The MSX-induced nuclear localization of GLN3 required the TOR-controlled, type 2A-related phosphatase SIT4. Other TOR-controlled transcription factors, GAT1/NIL1, MSN2, MSN4, and an unknown factor involved in the expression of ribosomal protein genes, were not affected by glutamine starvation. These findings suggest that the TOR pathway senses glutamine. Furthermore, as glutamine starvation affects only a subset of TOR-controlled transcription factors, TOR appears to discriminate between different nutrient conditions to elicit a response appropriate to a given condition.
MeSH Terms
Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
Carrier Proteins/genetics
Cation Transport Proteins
DNA-Binding Proteins/metabolism
Enzyme Inhibitors/pharmacology
Fungal Proteins/genetics,metabolism
Gene Expression
Glutamate-Ammonia Ligase/antagonists & inhibitors
Glutamic Acid/metabolism
Glutamine/metabolism
Intracellular Fluid/metabolism
Methionine Sulfoximine/pharmacology
Phosphatidylinositol 3-Kinases
Phosphotransferases (Alcohol Group Acceptor)/genetics,metabolism
Repressor Proteins
Saccharomyces cerevisiae/drug effects,growth & development,metabolism
Saccharomyces cerevisiae Proteins
Signal Transduction
Trans-Activators/metabolism
Transcription Factors
Chemicals
Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
Carrier Proteins
Cation Transport Proteins
DNA-Binding Proteins
Enzyme Inhibitors
Fungal Proteins
GLN3 protein, S cerevisiae
MEP2 protein, S cerevisiae
RTG1 protein, S cerevisiae
RTG3 protein, S cerevisiae
Repressor Proteins
Saccharomyces cerevisiae Proteins
Trans-Activators
Transcription Factors
Glutamine
Methionine Sulfoximine
Glutamic Acid
Phosphotransferases (Alcohol Group Acceptor)
TOR1 protein, S cerevisiae
Glutamate-Ammonia Ligase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Crespo José L
Division of Biochemistry, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland.
Powers Ted
Fowler Brian
Hall Michael N
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