title: Interactomics of Qa-SNARE in Arabidopsis thaliana.
abstract: Membrane trafficking in plants is involved in cellular development and the adaptation to various environmental changes. SNARE (soluble N-ethylmaleimide-sensitive factor attachment receptor) proteins mediate the fusion between vesicles and organelles to facilitate transport cargo proteins in cells. To characterize further the SNARE protein networks in cells, we carried out interactome analysis of SNARE proteins using 12 transgenic Arabidopsis thaliana plants expressing green fluorescent protein (GFP)-tagged Qa-SNAREs (SYP111, SYP121, SYP122, SYP123, SYP132, SYP21, SYP22, SYP31, SYP32, SYP41, SYP42 and SYP43). Microsomal fractions were prepared from each transgenic root, and subjected to immunoprecipitation (IP) using micromagnetic beads coupled to anti-GFP antibodies. To identify Qa-SNARE-interacting proteins, all immunoprecipitated products were then subjected to mass spectrometric (IP-MS) analysis. The IP-MS data revealed not only known interactions of SNARE proteins, but also unknown interactions. The IP-MS results were next categorized by gene ontology analysis. The data revealed that categories of cellular component organization, the cytoskeleton and endosome were enriched in the SYP2, SYP3 and SYP4 groups. In contrast, transporter activity was classified specifically in the SYP132 group. We also identified a novel interaction between SYP22 and VAMP711, which was validated using co-localization analysis with confocal microscopy and IP. Additional novel SNARE-interacting proteins play roles in vesicle transport and lignin biosynthesis, and were identified as membrane microdomain-related proteins. We propose that Qa-SNARE interactomics is useful for understanding SNARE interactions across the whole cell.
authors.affiliation: Plant Global Educational Project, Nara Institute of Science and Technology, Ikoma, 630-0192 Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Fujiwara
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Uemura
authors.forename: Kazuo
authors.initials: K
authors.lastname: Ebine
authors.forename: Yuka
authors.initials: Y
authors.lastname: Nishimori
authors.forename: Takashi
authors.initials: T
authors.lastname: Ueda
authors.forename: Akihiko
authors.initials: A
authors.lastname: Nakano
authors.forename: Masa H
authors.initials: MH
authors.lastname: Sato
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Fukao
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D047468:Immunoprecipitation; D058977:Molecular Sequence Annotation; D030821:Plants, Genetically Modified; D011485:Protein Binding; D025941:Protein Interaction Mapping; D021381:Protein Transport; D050765:Qa-SNARE Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D050765:Qa-SNARE Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins
keywords: Endomembrane; Immunoprecipitation; Interactomics; Mass spectrometry; SNARE