Interactome of the plant-specific ESCRT-III component AtVPS2.2 in Arabidopsis thaliana

Verena Ibl, Edina Csaszar, Nicole Schlager, Susanne Neubert, Christoph Spitzer, Marie-Theres Hauser

    Publications: Contribution to journalArticlePeer Reviewed

    Abstract

    The endosomal sorting complexes required for transport (ESCRT) guides transmembrane proteins to domains that bud away from the cytoplasm. The ESCRT machinery consists of four complexes. ESCRT complexes 0-II are important for cargo recognition and concentration via ubiquitin binding. Most of the membrane bending function is mediated by the large multimeric ESCRT-III complex and associated proteins. Here we present the first in vivo proteome analysis of a member of the ESCRT-III complex which is unique to the plant kingdom. We show with LC-MS/MS, yeast-two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) that coimmunoprecipitated proteins from Arabidopsis thaliana roots expressing a functional GFP-tagged VACUOLAR PROTEIN SORTING 2.2 (AtVPS2.2) protein are members of the ESCRT-III complex and associated proteins. Therefore we propose that at least in plants the large ESCRT-III membrane scaffolding complex consists of a mixture of SNF7, VPS2 and the associated VPS46 and VPS60 proteins. Apart from transmembrane proteins, numerous membrane-associated but also nuclear and extracellular proteins have been identified, indicating that AtVPS2.2 might be involved in processes beyond the classical ESCRT role. This study is the first in vivo proteome analysis with a tagged ESCRT-III component demonstrating the feasibility of this approach and provides numerous starting points for the investigation of the biological process in which AtVPS2.2 is involved.

    Original languageEnglish
    Pages (from-to)397-411
    Number of pages15
    JournalJournal of Proteome Research
    Volume11
    Issue number1
    DOIs
    Publication statusPublished - 1 Jan 2012

    Austrian Fields of Science 2012

    • 106052 Cell biology
    • 106023 Molecular biology

    Keywords

    • Arabidopsis
    • Arabidopsis Proteins
    • Cell Membrane Structures
    • Cell Nucleus
    • Dynamins
    • Endosomal Sorting Complexes Required for Transport
    • Green Fluorescent Proteins
    • Immunoprecipitation
    • Phospholipase D
    • Plant Roots
    • Protein Binding
    • Protein Transport
    • Proteome
    • Recombinant Fusion Proteins
    • Two-Hybrid System Techniques
    • Vesicular Transport Proteins
    • Journal Article
    • Research Support, Non-U.S. Gov't

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