title: The Qb-SNARE Memb11 interacts specifically with Arf1 in the Golgi apparatus of Arabidopsis thaliana.
abstract: The SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins are critical for the function of the secretory pathway. The SNARE Memb11 is involved in membrane trafficking at the ER-Golgi interface. The aim of the work was to decipher molecular mechanisms acting in Memb11-mediated ER-Golgi traffic. In mammalian cells, the orthologue of Memb11 (membrin) is potentially involved in the recruitment of the GTPase Arf1 at the Golgi membrane. However molecular mechanisms associated to Memb11 remain unknown in plants. Memb11 was detected mainly at the cis-Golgi and co-immunoprecipitated with Arf1, suggesting that Arf1 may interact with Memb11. This interaction of Memb11 with Arf1 at the Golgi was confirmed by in vivo BiFC (Bimolecular Fluorescence Complementation) experiments. This interaction was found to be specific to Memb11 as compared to either Memb12 or Sec22. Using a structural bioinformatic approach, several sequences in the N-ter part of Memb11 were hypothesized to be critical for this interaction and were tested by BiFC on corresponding mutants. Finally, by using both in vitro and in vivo approaches, we determined that only the GDP-bound form of Arf1 interacts with Memb11. Together, our results indicate that Memb11 interacts with the GDP-bound form of Arf1 in the Golgi apparatus.
authors.affiliation: CNRS-University of Bordeaux, UMR 5200 Membrane Biogenesis Laboratory, INRA Bordeaux Aquitaine, 33140 Villenave d'Ornon, France.
authors.forename: Claireline
authors.initials: C
authors.lastname: Marais
authors.affiliation: CNRS-University of Bordeaux, UMR 5200 Membrane Biogenesis Laboratory, INRA Bordeaux Aquitaine, 33140 Villenave d'Ornon, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Wattelet-Boyer
authors.affiliation: CNRS-University of Bordeaux, UMR 5200 Membrane Biogenesis Laboratory, INRA Bordeaux Aquitaine, 33140 Villenave d'Ornon, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Bouyssou
authors.affiliation: University of Bordeaux- INP Bordeaux, BPRVS, EA4135, F-33000 Bordeaux, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Hocquellet
authors.affiliation: Proteome platform, Functional Genomic Center of Bordeaux, University of Bordeaux, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France.
authors.forename: Jean-William
authors.initials: JW
authors.lastname: Dupuy
authors.affiliation: Bordeaux Imaging Center, UMS 3420 CNRS, US4 INSERM, University of Bordeaux, 33000 Bordeaux, France.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Batailler
authors.affiliation: Bordeaux Imaging Center, UMS 3420 CNRS, US4 INSERM, University of Bordeaux, 33000 Bordeaux, France.
authors.forename: Lysiane
authors.initials: L
authors.lastname: Brocard
authors.affiliation: CNRS-University of Bordeaux, UMR 5200 Membrane Biogenesis Laboratory, INRA Bordeaux Aquitaine, 33140 Villenave d'Ornon, France.
authors.forename: Yohann
authors.initials: Y
authors.lastname: Boutté
authors.affiliation: CNRS-University of Bordeaux, UMR 5200 Membrane Biogenesis Laboratory, INRA Bordeaux Aquitaine, 33140 Villenave d'Ornon, France.
authors.forename: Lilly
authors.initials: L
authors.lastname: Maneta-Peyret
authors.affiliation: CNRS-University of Bordeaux, UMR 5200 Membrane Biogenesis Laboratory, INRA Bordeaux Aquitaine, 33140 Villenave d'Ornon, France Bordeaux Imaging Center, UMS 3420 CNRS, US4 INSERM, University of Bordeaux, 33000 Bordeaux, France patrick.moreau@u-bordeaux.fr.
authors.forename: Patrick
authors.initials: P
authors.lastname: Moreau
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism*; D050766:Qb-SNARE Proteins / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: C503841:ARF1 protein, Arabidopsis; D029681:Arabidopsis Proteins; D004268:DNA-Binding Proteins; C000609725:Memb11 protein, Arabidopsis; D050766:Qb-SNARE Proteins; D014157:Transcription Factors
keywords: Arf1; GTPase; Golgi; Memb11; SNARE.; protein interaction; secretory pathway