Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex
| Autor: | Christoph, Kluge, Thorsten, Seidel, Susanne, Bolte, Shanti S, Sharma, Miriam, Hanitzsch, Beatrice, Satiat-Jeunemaitre, Joachim, Ross, Markus, Sauer, Dortje, Golldack, Karl-Josef, Dietz |
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| Přispěvatelé: | Biochemistry and Physiology of Plants, Universität Bielefeld = Bielefeld University, Institut des sciences du végétal (ISV), Centre National de la Recherche Scientifique (CNRS), Department of Biosciences, H. P. University, Applied Laser Physics and Laser Spectroscopy |
| Jazyk: | angličtina |
| Rok vydání: | 2004 |
| Předmět: |
MESH: Fluorescence Resonance Energy Transfer
MESH: Sequence Homology Amino Acid MESH: Trans Arabidopsis MESH: Onions MESH: Plant Roots Caryophyllaceae MESH: Amino Acid Sequence MESH: Protein Isoforms Endoplasmic Reticulum Plant Roots Polymerase Chain Reaction Epitopes MESH: Protein Structure Tertiary Onions Fluorescence Resonance Energy Transfer Protein Isoforms MESH: Arabidopsis health care economics and organizations Plant Proteins MESH: Plant Proteins lcsh:Cytology Protoplasts MESH: Protoplasts MESH: Protein Subunits Immunohistochemistry MESH: Saccharomyces cerevisiae MESH: Plant Leaves MESH: Membrane Proteins Subcellular Fractions Research Article Vacuolar Proton-Translocating ATPases DNA Complementary MESH: Epitopes Recombinant Fusion Proteins Molecular Sequence Data MESH: Sequence Alignment Saccharomyces cerevisiae Transfection Zea mays MESH: Endoplasmic Reticulum MESH: Recombinant Fusion Proteins [SDV.BV]Life Sciences [q-bio]/Vegetal Biology Amino Acid Sequence lcsh:QH573-671 MESH: Molecular Sequence Data Sequence Homology Amino Acid Membrane Proteins MESH: Polymerase Chain Reaction MESH: Immunohistochemistry MESH: DNA Complementary Protein Structure Tertiary Plant Leaves Protein Subunits MESH: Subcellular Fractions MESH: Caryophyllaceae Sequence Alignment |
| Zdroj: | BMC Cell Biology, Vol 5, Iss 1, p 29 (2004) BMC Cell Biology BMC Cell Biology, BioMed Central, 2004, 5, pp.29. ⟨10.1186/1471-2121-5-29⟩ |
| ISSN: | 1471-2121 |
| DOI: | 10.1186/1471-2121-5-29⟩ |
| Popis: | Background Vacuolar H+-ATPases are large protein complexes of more than 700 kDa that acidify endomembrane compartments and are part of the secretory system of eukaryotic cells. They are built from 14 different (VHA)-subunits. The paper addresses the question of sub-cellular localisation and subunit composition of plant V-ATPase in vivo and in vitro mainly by using colocalization and fluorescence resonance energy transfer techniques (FRET). Focus is placed on the examination and function of the 95 kDa membrane spanning subunit VHA-a. Showing similarities to the already described Vph1 and Stv1 vacuolar ATPase subunits from yeast, VHA-a revealed a bipartite structure with (i) a less conserved cytoplasmically orientated N-terminus and (ii) a membrane-spanning C-terminus with a higher extent of conservation including all amino acids shown to be essential for proton translocation in the yeast. On the basis of sequence data VHA-a appears to be an essential structural and functional element of V-ATPase, although previously a sole function in assembly has been proposed. Results To elucidate the presence and function of VHA-a in the plant complex, three approaches were undertaken: (i) co-immunoprecipitation with antibodies directed to epitopes in the N- and C-terminal part of VHA-a, respectively, (ii) immunocytochemistry approach including co-localisation studies with known plant endomembrane markers, and (iii) in vivo-FRET between subunits fused to variants of green fluorescence protein (CFP, YFP) in transfected cells. Conclusions All three sets of results show that V-ATPase contains VHA-a protein that interacts in a specific manner with other subunits. The genomes of plants encode three genes of the 95 kDa subunit (VHA-a) of the vacuolar type H+-ATPase. Immuno-localisation of VHA-a shows that the recognized subunit is exclusively located on the endoplasmic reticulum. This result is in agreement with the hypothesis that the different isoforms of VHA-a may localize on distinct endomembrane compartments, as it was shown for its yeast counterpart Vph1. |
| Databáze: | OpenAIRE |
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