Abstract
LST8, a Saccharomyces cerevisiae gene encoding a 34-kD WD-repeat protein, was identified by mutations that caused defects in sorting Gap1p to the plasma membrane. Here, we report that the Gap1p sorting defect in the lst8-1 mutant results from derepression of Rtg1/3p activity and the subsequent accumulation of high levels of intracellular amino acids, which signal Gap1p sorting to the vacuole. To identify the essential function of Lst8p, we isolated lst8 mutants that are temperature-sensitive for growth. These mutants show hypersensitivity to rapamycin and derepressed Gln3p activity like cells with compromised TOR pathway activity. Like tor2 mutants, lst8 mutants also have cell wall integrity defects. Confirming a role for Lst8p in the TOR pathway, we find that Lst8p associates with both Tor1p and Tor2p and is a peripheral membrane protein that localizes to endosomal or Golgi membranes and cofractionates with Tor1p. Further, we show that a sublethal concentration of rapamycin mimics the Gap1p sorting defect of an lst8 mutant. Finally, the different effects of lst8 alleles on the activation of either the Rtg1/3p or Gln3p transcription factors reveal that these two pathways constitute distinct, genetically separable outputs of the Tor-Lst8 regulatory complex.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Transport Systems*
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Amino Acids / biosynthesis*
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Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
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Carrier Proteins / genetics
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Carrier Proteins / metabolism*
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Cell Cycle Proteins
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Cell Division / genetics
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Cell Membrane / genetics
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Cell Membrane / metabolism*
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Cells, Cultured
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Feedback, Physiological / genetics
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Gene Expression Regulation, Fungal / genetics
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Intracellular Signaling Peptides and Proteins*
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Mutation / genetics
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Phosphatidylinositol 3-Kinases / genetics
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Phosphatidylinositol 3-Kinases / metabolism*
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Phosphotransferases (Alcohol Group Acceptor) / genetics
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Phosphotransferases (Alcohol Group Acceptor) / metabolism*
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Protein Transport / genetics*
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Repressor Proteins / genetics
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Repressor Proteins / metabolism
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Saccharomyces cerevisiae / metabolism*
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism*
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Signal Transduction / genetics
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Temperature
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Vacuoles / genetics
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Vacuoles / metabolism
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ras GTPase-Activating Proteins / genetics
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ras GTPase-Activating Proteins / metabolism*
Substances
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Amino Acid Transport Systems
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Amino Acids
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Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
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Carrier Proteins
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Cell Cycle Proteins
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GAP1 protein, S cerevisiae
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GLN3 protein, S cerevisiae
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Intracellular Signaling Peptides and Proteins
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LST8 protein, S cerevisiae
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RTG1 protein, S cerevisiae
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Repressor Proteins
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Saccharomyces cerevisiae Proteins
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Transcription Factors
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ras GTPase-Activating Proteins
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Phosphotransferases (Alcohol Group Acceptor)
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TOR1 protein, S cerevisiae
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TOR2 protein, S cerevisiae