Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex

Kluge C, Seidel T, Bolte S, Sharma SS, Hanitzsch M, Satiat-Jeunemaitre B, Ross J, Sauer M, Golldack D, Dietz K-J (2004)
BMC Cell Biology 5(1): 29.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
Kluge, C; Seidel, ThorstenUniBi; Bolte, S; Sharma, S. S.; Hanitzsch, M; Satiat-Jeunemaitre, B; Ross, J; Sauer, MarkusUniBi; Golldack, DortjeUniBi; Dietz, Karl-JosefUniBi
Abstract / Bemerkung
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.
BMC Cell Biology
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Kluge C, Seidel T, Bolte S, et al. Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex. BMC Cell Biology. 2004;5(1):29.
Kluge, C., Seidel, T., Bolte, S., Sharma, S. S., Hanitzsch, M., Satiat-Jeunemaitre, B., Ross, J., et al. (2004). Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex. BMC Cell Biology, 5(1), 29.
Kluge, C., Seidel, T., Bolte, S., Sharma, S. S., Hanitzsch, M., Satiat-Jeunemaitre, B., Ross, J., Sauer, M., Golldack, D., and Dietz, K. - J. (2004). Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex. BMC Cell Biology 5, 29.
Kluge, C., et al., 2004. Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex. BMC Cell Biology, 5(1), p 29.
C. Kluge, et al., “Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex”, BMC Cell Biology, vol. 5, 2004, pp. 29.
Kluge, C., Seidel, T., Bolte, S., Sharma, S.S., Hanitzsch, M., Satiat-Jeunemaitre, B., Ross, J., Sauer, M., Golldack, D., Dietz, K.-J.: Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex. BMC Cell Biology. 5, 29 (2004).
Kluge, C, Seidel, Thorsten, Bolte, S, Sharma, S. S., Hanitzsch, M, Satiat-Jeunemaitre, B, Ross, J, Sauer, Markus, Golldack, Dortje, and Dietz, Karl-Josef. “Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex”. BMC Cell Biology 5.1 (2004): 29.
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