Abstract
Each of the three plasma membrane Ato proteins is involved in ammonium signalling and the development of yeast colonies. This suggests that although these proteins are homologous, they do not functionally substitute for each other, but may form a functional complex. Here, we present a detailed combined FRET, FLIM and photobleaching study, which enabled us to detect interactions between Ato proteins found in distinct compartments of yeast cells. We thus show that the proteins Ato1p and Ato2p interact and can form complexes when present in the plasma membrane. No interaction was detected between Ato1p and Ato3p or Ato2p and Ato3p. In addition, using specially prepared strains, we were able to detect an interaction between molecules of the same Ato protein, namely Ato1p-Ato1p and Ato3p-Ato3p, but not Ato2p-Ato2p.
Copyright © 2012 Elsevier B.V. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cell Membrane / metabolism*
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Fluorescence Recovery After Photobleaching
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Fluorescence Resonance Energy Transfer / methods
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Gene Expression Regulation, Fungal
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Green Fluorescent Proteins / metabolism
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Membrane Proteins / metabolism
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Membrane Proteins / physiology*
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Membrane Transport Proteins / metabolism
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Membrane Transport Proteins / physiology*
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Models, Biological
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Protein Interaction Mapping
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Saccharomyces cerevisiae / chemistry*
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / metabolism
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Saccharomyces cerevisiae Proteins / physiology*
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Signal Transduction
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Spectrometry, Fluorescence / methods*
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Subcellular Fractions / metabolism
Substances
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ATO1 protein, S cerevisiae
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ATO2 protein, S cerevisiae
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ATO3 protein, S cerevisiae
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Membrane Proteins
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Membrane Transport Proteins
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Saccharomyces cerevisiae Proteins
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Green Fluorescent Proteins