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Reference - PMID:26689777 - Tor Signaling Regulates Transcription of Amino Acid Permeases through a GATA Transcription Factor Gaf1 in Fission Yeast.

Reference summary

PubMed ID
PMID:26689777
Title
Tor Signaling Regulates Transcription of Amino Acid Permeases through a GATA Transcription Factor Gaf1 in Fission Yeast.
Authors
Ma Y, Ma N, Liu Q, Qi Y, Manabe R, Furuyashiki T
Citation
PLoS One 2015;10(12):e0144677
Publication year
2015
Abstract
In the fission yeast, two Tor isoforms, Tor1 and Tor2, oppositely regulate gene expression of amino acid permeases. To elucidate the transcriptional machinery for these regulations, here we have employed the cap analysis of gene expression (CAGE), a method of analyzing expression profiles and identifying transcriptional start sites (TSSs). The loss of Tor1 decreased, and Tor2 inhibition by its temperature sensitive mutation increased, mRNA expression of isp5+, per1+, put4+ and SPBPB2B2.01. In contrast, the loss of Tor1 increased, and Tor2 inhibition decreased, the expression of cat1+. These changes were confirmed by semi-quantitative RT-PCR. These opposite effects by the loss of Tor1 and Tor2 inhibition appeared to occur evenly across multiple TSSs for the respective genes. The motif discovery analysis based on the CAGE results identified the GATA motifs as a potential cis-regulatory element for Tor-mediated regulation. In the luciferase reporter assay, the loss of Tor1 reduced, and Tor2 inhibition and nitrogen depletion increased, the activity of isp5+ promoter as well as that of a GATAAG reporter. One of the GATAAG motifs in isp5+ promoter was critical for its transcriptional activity, and a GATA transcription factor Gaf1 was critical for the activities of isp5+ promoter and the GATAAG reporter. Furthermore, Tor2 inhibition and nitrogen depletion induced nuclear localization of Gaf1 from the cytosol and its dephosphorylation. These results suggest that Tor2 inhibition, which is known to be induced by nitrogen depletion, promotes nuclear localization of Gaf1, thereby inducing isp5+ transcription through Gaf1 binding to the GATAAG motif in its promoter. Since Gaf1 was also critical for transcription of per1+ and put4+, Tor-Gaf1 signaling may coordinate transcription of multiple amino acid permeases according to nutrient availability.

Annotation

GO biological process

GO:1904262 - negative regulation of TORC1 signaling

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GO cellular component

GO:0000329 - fungal-type vacuole membrane

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Multi-locus phenotype

FYPO:0004457 - decreased protein localization to nucleus during nitrogen starvation

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FYPO:0001117 - decreased RNA level during vegetative growth

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FYPO:0001355 - decreased vegetative cell population growth

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FYPO:0000825 - increased RNA level during vegetative growth

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FYPO:0001357 - normal vegetative cell population growth

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FYPO:0001029 - resistance to canavanine

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Single locus phenotype

FYPO:0006148 - abolished transcription during nitrogen starvation

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FYPO:0001324 - decreased protein level during vegetative growth

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FYPO:0010009 - decreased protein localization to cell surface, with protein mislocalized to cytoplasm

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FYPO:0004457 - decreased protein localization to nucleus during nitrogen starvation

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FYPO:0006233 - decreased protein localization to vacuolar membrane

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FYPO:0001117 - decreased RNA level during vegetative growth

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FYPO:0001355 - decreased vegetative cell population growth

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FYPO:0002681 - increased protein phosphorylation during nitrogen starvation

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FYPO:0000825 - increased RNA level during vegetative growth

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FYPO:0001545 - normal growth on L-canavanine

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FYPO:0008372 - normal protein localization to cell surface during vegetative growth

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Genotypes:

FYPO:0001357 - normal vegetative cell population growth

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Genotypes:

FYPO:0001029 - resistance to canavanine

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