%0 Journal Article %1 Seidel2008 %A Seidel, Thorsten %A Schnitzer, Daniel %A Golldack, Dortje %A Sauer, Markus %A Dietz, Karl-Josef %C CURRENT SCIENCE GROUP, MIDDLESEX HOUSE, 34-42 CLEVELAND ST, LONDON W1T 4LB, ENGLAND %D 2008 %I BIOMED CENTRAL LTD %J BMC CELL BIOLOGY %K sauer %T Organelle-specific isoenzymes of plant V-ATPase as revealed by in vivo-FRET analysis %U http://dx.doi.org/10.1186/1471-2121-9-28 %V 9 %X Background: The V-ATPase ( VHA) is a protein complex of 13 different VHA-subunits. It functions as an ATP driven rotary-motor that electrogenically translocates H+ into endomembrane compartments. In Arabidopsis thaliana V-ATPase is encoded by 23 genes posing the question of specific versus redundant function of multigene encoded isoforms. Results: The transmembrane topology and stoichiometry of the proteolipid VHA-c'' as well as the stoichiometry of the membrane integral subunit VHA-e within the V-ATPase complex were investigated by in vivo fluorescence resonance energy transfer ( FRET). VHA-c'', VHA-e1 and VHA-e2, VHA-a, VHA-c3, truncated variants of VHA-c3 and a chimeric VHA-c/VHA-c'' hybrid were fused to cyan ( CFP) and yellow fluorescent protein ( YFP), respectively. The constructs were employed for transfection experiments with Arabidopsis thaliana mesophyll protoplasts. Subcellular localization and FRET analysis by confocal laser scanning microscopy ( CLSM) demonstrated that (i.) the N- and C-termini of VHA-c'' are localised in the vacuolar lumen, ( ii.) one copy of VHA-c'' is present within the VHA-complex, and ( iii.) VHA-c'' is localised at the ER and associated Golgi bodies. ( iv.) A similar localisation was observed for VHA-e2, whereas ( v.) the subcellular localisation of VHA-e1 indicated the trans Golgi network ( TGN)-specifity of this subunit. Conclusion: The plant proteolipid ring is a highly flexible protein subcomplex, tolerating the incorporation of truncated and hybrid proteolipid subunits, respectively. Whereas the membrane integral subunit VHA-e is present in two copies within the complex, the proteolipid subunit VHA-c'' takes part in complex formation with only one copy. However, neither VHA-c'' isoform 1 nor any of the two VHA-e isoforms were identified at the tonoplast. This suggest a function in endomembrane specific VHA-assembly or targeting rather than proton transport.

@article{Seidel2008, abstract = {Background: The V-ATPase ( VHA) is a protein complex of 13 different VHA-subunits. It functions as an ATP driven rotary-motor that electrogenically translocates H+ into endomembrane compartments. In Arabidopsis thaliana V-ATPase is encoded by 23 genes posing the question of specific versus redundant function of multigene encoded isoforms. Results: The transmembrane topology and stoichiometry of the proteolipid VHA-c{''} as well as the stoichiometry of the membrane integral subunit VHA-e within the V-ATPase complex were investigated by in vivo fluorescence resonance energy transfer ( FRET). VHA-c{''}, VHA-e1 and VHA-e2, VHA-a, VHA-c3, truncated variants of VHA-c3 and a chimeric VHA-c/VHA-c{''} hybrid were fused to cyan ( CFP) and yellow fluorescent protein ( YFP), respectively. The constructs were employed for transfection experiments with Arabidopsis thaliana mesophyll protoplasts. Subcellular localization and FRET analysis by confocal laser scanning microscopy ( CLSM) demonstrated that (i.) the N- and C-termini of VHA-c{''} are localised in the vacuolar lumen, ( ii.) one copy of VHA-c{''} is present within the VHA-complex, and ( iii.) VHA-c{''} is localised at the ER and associated Golgi bodies. ( iv.) A similar localisation was observed for VHA-e2, whereas ( v.) the subcellular localisation of VHA-e1 indicated the trans Golgi network ( TGN)-specifity of this subunit. Conclusion: The plant proteolipid ring is a highly flexible protein subcomplex, tolerating the incorporation of truncated and hybrid proteolipid subunits, respectively. Whereas the membrane integral subunit VHA-e is present in two copies within the complex, the proteolipid subunit VHA-c{''} takes part in complex formation with only one copy. However, neither VHA-c{''} isoform 1 nor any of the two VHA-e isoforms were identified at the tonoplast. This suggest a function in endomembrane specific VHA-assembly or targeting rather than proton transport.}, added-at = {2011-03-01T10:57:13.000+0100}, address = {CURRENT SCIENCE GROUP, MIDDLESEX HOUSE, 34-42 CLEVELAND ST, LONDON W1T 4LB, ENGLAND}, affiliation = {Seidel, T (Reprint Author), Univ Bielefeld, Dept Biochem \& Physiol Plants, W5, D-33501 Bielefeld, Germany. {[}Seidel, Thorsten; Schnitzer, Daniel; Golldack, Dortje; Dietz, Karl-Josef] Univ Bielefeld, Dept Biochem \& Physiol Plants, D-33501 Bielefeld, Germany. {[}Sauer, Markus] Univ Bielefeld, Dept Laser Phys, D-33501 Bielefeld, Germany.}, article-number = {28}, author = {Seidel, Thorsten and Schnitzer, Daniel and Golldack, Dortje and Sauer, Markus and Dietz, Karl-Josef}, author-email = {thorsten.seidel@uni-bielefeld.de daniel.schnitzer@uni-bielefeld.de dortje.golldack@uni-bielefeld.de markus.sauer@physik.uni-bielefeld.de karl-josef.dietz@uni-bielefeld.de}, biburl = {https://www.bibsonomy.org/bibtex/22bccc8c0ce2276003ef8f552fa5377dc/reichert}, cited-references = {ARECHAGA I, 2001, FEBS LETT, V494, P1. AVIEZARHAGAI K, 2004, J EXP BIOL, V206, P3227. BOEVINK P, 1998, PLANT J, V15, P441. CARTER C, 2004, PLANT CELL, V16, P3285. CHARSKY CMH, 2000, J BIOL CHEM, V275, P37232. COMPTON MA, 2006, J BIOL CHEM, V281, P15312, DOI 10.1074/jbc.M600890200. CROSS RL, 2004, FEBS LETT, V576, P1, DOI 10.1016/j.febslet.2004.08.065. CURTIS KK, 2002, J BIOL CHEM, V277, P2716. DETTMER J, 2005, PLANT J, V41, P117, DOI 10.1111/j.1365-313X.2004.02282.x. DETTMER J, 2006, PLANT CELL, V18, P715, DOI 10.1105/tpc.105.037978. DOMGALL I, 2002, J BIOL CHEM, V277, P13115. DROBNY M, 2002, BBA-BIOMEMBRANES, V1564, P243. DU J, 2006, J CELL SCI, V119, P2542, DOI 10.1242/jcs.02983. FLANNERY AR, 2004, J BIOL CHEM, V279, P39856, DOI 10.1074/jbc.M406767200. GIBSON LCD, 2002, BIOCHEM J 3, V366, P911, DOI 10.1042/BJ20020171. GRABE M, 2000, BIOPHYS J, V78, P2798. GRAHAM LA, 1998, J CELL BIOL, V142, P39. GRAHAM LA, 2003, J BIOENERG BIOMEMBR, V35, P301. GRUBER G, 2001, J EXP BIOL, V204, P2597. HARRISON M, 2003, J BIOENERG BIOMEMBR, V35, P337. HILL KJ, 1994, MOL BIOL CELL, V5, P1039. HIRATA R, 1997, J BIOL CHEM, V272, P4795. HIRATA T, 2003, J BIOL CHEM, V278, P23714, DOI 10.1074/jbc.M302756200. KADER MA, 2006, J EXP BOT, V57, P4257, DOI 10.1093/jxb/erl199. KANE PM, 1999, J BIOL CHEM, V274, P17275. KAWASAKINISHI S, 2003, FEBS LETT, V545, P76, DOI 10.1016/S0014-5793(03)00396-X. KLUGE C, 2004, BMC CELL BIOL, V5. LUDWIG J, 1998, J BIOL CHEM, V273, P10939. LUTTGE U, 2001, CELL MOL BIOL LETT, V6, P356. MALKUS P, 2004, MOL BIOL CELL, V15, P5075, DOI 10.1091/mbc.E04-06-0514. MARIAUX JB, 1997, PROTOPLASMA, V196, P181. MATSUOKA K, 1997, PLANT CELL, V9, P533. NISHI T, 2001, J BIOL CHEM, V276, P34122. NISHI T, 2003, J BIOL CHEM, V278, P5821, DOI 10.1074/jbc.M209875200. PADMANABAN S, 2004, PLANT PHYSIOL, V134, P1514, DOI 10.1104/pp.103.034025. PETERS C, 2001, NATURE, V409, P581. POWELL B, 2000, J BIOL CHEM, V275, P23654. QI J, 2007, J BIOL CHEM, V282, P24743. RATAJCZAK R, 2000, BBA-BIOMEMBRANES, V1465, P17. SAMBADE M, 2004, J BIOL CHEM, V279, P17361, DOI 10.1074/jbc.M314104200. SCHUMACHER K, 1999, GENE DEV, V13, P3259. SEIDEL T, 2004, J BIOTECHNOL, V112, P165, DOI 10.1016/j.jbiotec.2004.04.027. SEIDEL T, 2005, FEBS LETT, V579, P4374, DOI 10.1016/j.febslet.2005.06.077. SHIMAOKA T, 2004, PLANT CELL PHYSIOL, V45, P672. STROMPEN G, 2005, PLANT J, V41, P125, DOI 10.1111/j.1365-313X.2004.02283.x. SZE H, 1999, PLANT CELL, V11, P677. SZE H, 2002, TRENDS PLANT SCI, V7, P157. URADE R, 2007, FEBS J, V274, P1152, DOI 10.1111/j.1742-4658.2007.05664.x. WANG Y, 2007, J BIOL CHEM, V282, P34058, DOI 10.1074/jbc.M704331200. WANG YR, 2004, J BIOL CHEM, V279, P44628, DOI 10.1074/jbc.M407345200.}, doc-delivery-number = {316AY}, groups = {public}, interhash = {8d3d80e9add28c77a0bf89a183b24a1f}, intrahash = {2bccc8c0ce2276003ef8f552fa5377dc}, issn = {1471-2121}, journal = {BMC CELL BIOLOGY}, journal-iso = {BMC Cell Biol.}, keywords = {sauer}, keywords-plus = {VACUOLAR H+-ATPASE; SUBUNIT C'' VMA16P; ENDOPLASMIC-RETICULUM; TRANSLOCATING ATPASE; SACCHAROMYCES-CEREVISIAE; ARABIDOPSIS-THALIANA; YEAST; MEMBRANE; COMPLEX; EXPRESSION}, language = {English}, month = {MAY 28}, number-of-cited-references = {50}, publisher = {BIOMED CENTRAL LTD}, subject-category = {Cell Biology}, times-cited = {2}, timestamp = {2011-03-09T14:16:20.000+0100}, title = {Organelle-specific isoenzymes of plant V-ATPase as revealed by in vivo-FRET analysis}, type = {Article}, unique-id = {ISI:000256922500001}, url = {http://dx.doi.org/10.1186/1471-2121-9-28}, username = {reichert}, volume = 9, year = 2008 }