title: Ato protein interactions in yeast plasma membrane revealed by fluorescence lifetime imaging (FLIM).
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.
authors.affiliation: Institute of Microbiology of the ASCR, Prague 4, Czech Republic.
authors.forename: Dita
authors.initials: D
authors.lastname: Strachotová
authors.forename: Aleš
authors.initials: A
authors.lastname: Holoubek
authors.forename: Helena
authors.initials: H
authors.lastname: Kučerová
authors.forename: Aleš
authors.initials: A
authors.lastname: Benda
authors.forename: Jana
authors.initials: J
authors.lastname: Humpolíčková
authors.forename: Libuše
authors.initials: L
authors.lastname: Váchová
authors.forename: Zdena
authors.initials: Z
authors.lastname: Palková
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D036681:Fluorescence Recovery After Photobleaching; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D015966:Gene Expression Regulation, Fungal; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology*; D026901:Membrane Transport Proteins / Q000378:metabolism / Q000502:physiology*; D008954:Models, Biological; D025941:Protein Interaction Mapping; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: C520069:ATO1 protein, S cerevisiae; C089361:ATO2 protein, S cerevisiae; C479828:ATO3 protein, S cerevisiae; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins