Neklesa TK~Davis RW, 2009

Pubmed ID 19521502
Title A genome-wide screen for regulators of TORC1 in response to amino acid starvation reveals a conserved Npr2/3 complex.
Authors Taavi K Neklesa, Ronald W Davis
Abstract TORC1 is a central regulator of cell growth in response to amino acid availability, yet little is known about how it is regulated. Here, we performed a reverse genetic screen in yeast for genes necessary to inactivate TORC1. The screen consisted of monitoring the expression of a TORC1 sensitive GFP-based transcriptional reporter in all yeast deletion strains using flow cytometry. We find that in response to amino acid starvation, but not to carbon starvation or rapamycin treatment, cells lacking NPR2 and NPR3 fail to fully (1) activate transcription factors Gln3/Gat1, (2) dephosphorylate TORC1 effector Npr1, and (3) repress ribosomal protein gene expression. Both mutants show proliferation defects only in media containing a low quality nitrogen source, such as proline or ammonia, whereas no defects are evident when cells are grown in the presence of glutamine or peptone mixture. Proliferation defects in npr2Delta and npr3Delta cells can be completely rescued by artificially inhibiting TORC1 by rapamycin, demonstrating that overactive TORC1 in both strains prevents their ability to adapt to an environment containing a low quality nitrogen source. A biochemical purification of each demonstrates that Npr2 and Npr3 form a heterodimer, and this interaction is evolutionarily conserved since the human homologs of NPR2 and NPR3 (NPRL2 and NPRL3, respectively) also co-immunoprecipitate. We conclude that, in yeast, the Npr2/3 complex mediates an amino acid starvation signal to TORC1.
Citation PLoS Genet. 2009; 5:e1000515


Download the list of datasets
Papers Phenotype Conditions Collection Tested mutants Data Details
Neklesa TK~Davis RW, 2009 TORC1 activity (Dal80pr-GFP) YPD [standard] hom ~5,100 None
Neklesa TK~Davis RW, 2009 TORC1 activity (Dal80pr-GFP) sirolimus [20 ng/ml] hom ~5,100 None
Neklesa TK~Davis RW, 2009 TORC1 activity (Dal80pr-GFP) YNB without amino acids [0.17%] hom ~5,100 None

Curation history

Oct. 10, 2016 Tested strains to request.
Oct. 10, 2016 Data waiting for tested.
April 18, 2017 Tested strains requested.
June 23, 2017 Tested strains requested.
Aug. 28, 2017 Tested strains abandoned.
Aug. 28, 2017 Data to load.
Feb. 2, 2018 Data abandoned.