[en] Mitochondrial F1FO-ATP synthase of chlorophycean algae is a complex partially embedded in the inner mitochondrial membrane that is isolated as a highly stable dimer of 1600 kDa. It comprises 17 polypeptides, nine of which (subunits Asa1 to 9) are not present in classical mitochondrial ATP synthases and appear to be exclusive of the chlorophycean lineage. In particular, subunits Asa2, Asa4 and Asa7 seem to constitute a section of the peripheral stalk of the enzyme. Here, we over-expressed and purified subunits Asa2, Asa4 and Asa7 and the corresponding amino-terminal and carboxy-terminal halves of Asa4 and Asa7 in order to explore their interactions in vitro, using immunochemical techniques, blue native electrophoresis and affinity chromatography. Asa4 and Asa7 interact strongly, mainly through their carboxy-terminal halves. Asa2 interacts with both Asa7 and Asa4, and also with subunit ?? in the F1 sector. The three Asa proteins form an Asa2/Asa4/Asa7 subcomplex. The entire Asa7 and the carboxy-terminal half of Asa4 seem to be instrumental in the interaction with Asa2. Based on these results and on computer-generated structural models of the three subunits, we propose a model for the Asa2/Asa4/Asa7 subcomplex and for its disposition in the peripheral stalk of the algal ATP synthase.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Miranda Astudillo, Héctor Vicente ; Universidad Nacional Autónoma de México > Instituto de Fisiología Celular > Departamento de Genética Molecular
Cano-Estrada, Arceli; Universidad Nacional Autónoma de México > Instituto de Fisiología Celular > Departamento de Genética Molecular
Vázquez-Acevedo, Miriam; Universidad Nacional Autónoma de México > Instituto de Fisiología Celular > Departamento de Genética Molecular
Colina-Tenorio, Lilia; Universidad Nacional Autónoma de México > Instituto de Fisiología Celular > Departamento de Genética Molecular
Downie-Velasco, Angela; Universidad Nacional Autónoma de México > Instituto de Fisiología Celular > Departamento de Genética Molecular
Cardol, Pierre ; Université de Liège - ULiège > Département des sciences de la vie > Génétique
Remacle, Claire ; Université de Liège - ULiège > Département des sciences de la vie > Génétique
Domínguez-Ramírez, Lenin; Universidad Nacional Autónoma de México > Instituto de Fisiología Celular > Departamento de Genética Molecular
González-Halphen, Diego; Universidad Nacional Autónoma de México > Instituto de Fisiología Celular > Departamento de Genética Molecular
Language :
English
Title :
Interactions of subunits Asa2, Asa4 and Asa7 in the peripheral stalk of the mitochondrial ATP synthase of the chlorophycean alga Polytomella sp.
Devenish, R.J., Prescott, M., Rodgers, A.J., The structure and function of mitochondrial F1F0-ATP synthases (2008) Int. Rev. Cell Mol. Biol., 267, pp. 1-58
Seelert, H., Dencher, N.A., ATP synthase superassemblies in animals and plants: Two or more are better (2011) Biochim. Biophys. Acta, 1807, pp. 1185-1197
Junge, W., Sielaff, H., Engelbrecht, S., Torque generation and elastic power transmission in the rotary F(O)F(1)-ATPase (2009) Nature, 459, pp. 364-370
Van Lis, R., Atteia, A., Mendoza-Hernández, G., González-Halphen, D., Identification of novel mitochondrial protein components of Chlamydomonas reinhardtii. A proteomic approach (2003) Plant Physiol., 132, pp. 318-330
Dudkina, N.V., Heinemeyer, J., Keegstra, W., Boekema, E.J., Braun, H.P., Structure of dimeric ATP synthase from mitochondria: An angular association of monomers induces the strong curvature of the inner membrane (2005) FEBS Lett., 579, pp. 5769-5772
Dudkina, N.V., Sunderhaus, S., Braun, H.P., Boekema, E.J., Characterization of dimeric ATP synthase and cristae membrane ultrastructure from Saccharomyces and Polytomella mitochondria (2006) FEBS Lett., 580, pp. 3427-3432
Cano-Estrada, A., Vázquez-Acevedo, M., Villavicencio-Queijeiro, A., Figueroa-Martínez, F., Miranda-Astudillo, H., Cordeiro, Y., Mignaco, J.A., González-Halphen, D., Subunit-subunit interactions and overall topology of the dimeric mitochondrial ATP synthase of Polytomella sp (2010) Biochim. Biophys. Acta, 1797, pp. 1439-1448
Dudkina, N.V., Oostergetel, G.T., Lewejohann, D., Braun, H.P., Boekema, E.J., Row-like organization of ATP synthase in intact mitochondria determined by cryo-electron tomography (2010) Biochim. Biophys. Acta, 1797, pp. 272-277
Atteia, A., Dreyfus, G., González-Halphen, D., Characterization of the alpha and beta-subunits of the F0F1-ATPase from the alga Polytomella spp., a colorless relative of Chlamydomonas reinhardtii (1997) Biochim. Biophys. Acta, 1320, pp. 275-284
Vázquez-Acevedo, M., Cardol, P., Cano-Estrada, A., Lapaille, M., Remacle, C., González-Halphen, D., The mitochondrial ATP synthase of chlorophycean algae contains eight subunits of unknown origin involved in the formation of an atypical stator-stalk and in the dimerization of the complex (2006) J. Bioenerg. Biomembr., 38, pp. 271-282
Van Lis, R., Mendoza-Hernández, G., Groth, G., Atteia, A., New insights into the unique structure of the F0F1-ATP synthase from the chlamydomonad algae Polytomella sp. and Chlamydomonas reinhardtii (2007) Plant Physiol., 144, pp. 1190-1199
Villavicencio-Queijeiro, A., Vázquez-Acevedo, M., Cano-Estrada, A., Zarco-Zavala, M., Tuena De Gómez, M., Mignaco, J.A., Freire, M.M., González-Halphen, D., The fully-active and structurally-stable form of the mitochondrial ATP synthase of Polytomella sp. is dimeric (2009) J. Bioenerg. Biomembr., 41, pp. 1-13
Lapaille, M., Escobar-Ramírez, A., Degand, H., Baurain, D., Rodríguez-Salinas, E., Coosemans, N., Boutry, M., Cardol, P., Atypical subunit composition of the chlorophycean mitochondrial F 1FO-ATP synthase and role of Asa7 protein in stability and oligomycin resistance of the enzyme (2010) Mol. Biol. Evol., 27, pp. 1630-1644
Funes, S., Davidson, E., Claros, M.G., Van Lis, R., Pérez-Martínez, X., Vázquez-Acevedo, M., King, M.P., González-Halphen, D., The typically mitochondrial DNA-encoded ATP6 subunit of the F1F0-ATPase is encoded by a nuclear gene in Chlamydomonas reinhardtii (2002) J. Biol. Chem., 277, pp. 6051-6058
Cardol, P., González-Halphen, D., Reyes-Prieto, A., Baurain, D., Matagne, R.F., Remacle, C., The mitochondrial oxidative phosphorylation proteome of Chlamydomonas reinhardtii deduced from the Genome Sequencing Project (2005) Plant Physiol., 137, pp. 447-459
Gutiérrez-Cirlos, E.B., Antaramian, A., Vázquez-Acevedo, M., Coria, R., González-Halphen, D., A highly active ubiquinol-cytochrome c reductase (bc1 complex) from the colorless alga Polytomella spp., a close relative of Chlamydomonas. Characterization of the heme binding site of cytochrome c1 (1994) J. Biol. Chem., 269, pp. 9147-9154
Pérez-Martínez, X., Vázquez-Acevedo, M., Tolkunova, E., Funes, S., Claros, M.G., Davidson, E., King, M.P., González-Halphen, D., Unusual location of a mitochondrial gene. Subunit III of cytochrome C oxidase is encoded in the nucleus of chlamydomonad algae (2000) J. Biol. Chem., 275, pp. 30144-30152
Van Lis, R., González-Halphen, D., Atteia, A., Divergence of the mitochondrial electron transport chains from the green alga Chlamydomonas reinhardtii and its colorless close relative Polytomella sp (2005) Biochim. Biophys. Acta, 1708, pp. 23-34
Schägger, H., Denaturing electrophoretic techniques (1994) A Practical Guide to Membrane Protein Purification, pp. 59-79. , G. von Jagow, H. Schägger, Academic Press San Diego
Schägger, H., Native gel electrophoresis (1994) A Practical Guide to Membrane Protein Purification, pp. 81-104. , G. von Jagow, H. Schägger, Academic Press San Diego
Markwell, M.A.K., Hass, S.M., Biber, L.L., Tolbert, N.E., A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples (1978) Anal. Biochem., 87, pp. 206-210
Burgess, R.R., Refolding solubilized inclusion body proteins (2009) Methods Enzymol., 463, pp. 259-282
Schägger, H., Aquila, H., Von Jagow, G., Coomassie blue-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for direct visualization of polypeptides during electrophoresis (1988) Anal. Biochem., 173, pp. 201-205
Towbin, H., Staehelin, T., Gordon, J., Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications (1979) Proc. Natl. Acad. Sci. U. S. A., 76, pp. 4350-4354
González-Halphen, D., Lindorfer, M.A., Capaldi, R.A., Subunit arrangement in beef heart complex III (1988) Biochemistry, 27, pp. 7021-7031
Hall, R.A., Studying protein-protein interactions via blot overlay or Far Western blot (2004) Methods Mol. Biol., 261, pp. 167-174
Raman, S., Vernon, R., Thompson, J., Tyka, M., Sadreyev, R., Pei, J., Kim, D., Baker, D., Structure prediction for CASP8 with all-atom refinement using Rosetta (2009) Proteins, 77 (SUPPL. 9), pp. 89-99
Zhang, Y., I-TASSER server for protein 3D structure prediction (2008) BMC Bioinforma., 9, p. 40
Pettersen, E.F., Goddard, T.D., Huang, C.C., Couch, G.S., Greenblatt, D.M., Meng, E.C., Ferrin, T.E., UCSF chimera - A visualization system for exploratory research and analysis (2004) J. Comput. Chem., 25, pp. 1605-1612
Tovchigrechko, A., Vakser, I.A., GRAMM-X public web server for protein-protein docking (2006) Nucleic Acids Res., 34, pp. 310-W314
Wallace, A.C., Laskowski, R.A., Thornton, J.M., LIGPLOT: A program to generate schematic diagrams of protein-ligand interactions (1995) Protein Eng., 8, pp. 127-134
Dickson, V.K., Silvester, J.A., Fearnley, I.M., Leslie, A.G., Walker, J.E., On the structure of the stator of the mitochondrial ATP synthase (2006) EMBO J., 25, pp. 2911-2918
Poetsch, A., Berzborn, R.J., Heberle, J., Link, T.A., Dencher, N.A., Seelert, H., Biophysics and bioinformatics reveal structural differences of the two peripheral stalk subunits in chloroplast ATP synthase (2007) J. Biochem., 141, pp. 411-420
Welch, A.K., Bostwick, C.J., Cain, B.D., Manipulations in the peripheral stalk of the Saccharomyces cerevisiae F1F0-ATP synthase (2011) J. Biol. Chem., 286, pp. 10155-10162
Bi, Y., Watts, J.C., Bamford, P.K., Briere, L.K., Dunn, S.D., Probing the functional tolerance of the b subunit of Escherichia coli ATP synthase for sequence manipulation through a chimera approach (2008) Biochim. Biophys. Acta, 1777, pp. 583-591
Claggett, S.B., O'Neil Plancher, M., Dunn, S.D., Cain, B.D., The b subunits in the peripheral stalk of F1F0 ATP synthase preferentially adopt an offset relationship (2009) J. Biol. Chem., 284, pp. 16531-16540
Priya, R., Biukovic, G., Gayen, S., Vivekanandan, S., Grüber, G., Solution structure, determined by nuclear magnetic resonance, of the b30-82 domain of subunit b of Escherichia coli F1FO ATP synthase (2009) J. Bacteriol., 191, pp. 7538-7544
Balakrishna, A.M., Hunke, C., Grüber, G., The structure of subunit e of the Pyrococcus horikoshii OT3 A-ATP synthase gives insight into the elasticity of the peripheral stalk (2012) J. Mol. Biol., 420, pp. 155-163
Fariselli, P., Molinini, D., Casadio, R., Krogh, A., Prediction of structurally-determined coiled-coil domains with hidden Markov models (2007) Lect. Notes Comput. Sci, 4414, pp. 292-302
Walker, J.E., Dickson, V.K., The peripheral stalk of the mitochondrial ATP synthase (2006) Biochim. Biophys. Acta, 1757, pp. 286-296
McLachlin, D.T., Dunn, S.D., Disulfide linkage of the b and delta subunits does not affect the function of the Escherichia coli ATP synthase (2000) Biochemistry, 39, pp. 3486-3490
Vonck, J., Pisa, K.Y., Morgner, N., Brutschy, B., Müller, V., Three-dimensional structure of A1A0 ATP synthase from the hyperthermophilic archaeon Pyrococcus furiosus by electron microscopy (2009) J. Biol. Chem., 284, pp. 10110-10119
Rees, D.M., Leslie, A.G., Walker, J.E., The structure of the membrane extrinsic region of bovine ATP synthase (2009) Proc. Natl. Acad. Sci. U. S. A., 106, pp. 21597-21601
Balabaskaran, N.P., Dudkina, N.V., Kane, L.A., Van Eyk, J.E., Boekema, E.J., Mather, M.W., Vaidya, A.B., Highly divergent mitochondrial ATP synthase complexes in Tetrahymena thermophila (2010) PLoS Biol., 8, p. 1000418
Zikova, A., Schnaufer, A., Dalley, R.A., Panigrahi, A.K., Stuart, K.D., The F(0)F(1)-ATP synthase complex contains novel subunits and is essential for procyclic Trypanosoma brucei (2009) PLoS Pathog., 5, p. 1000436
Wächter, A., Bi, Y., Dunn, S.D., Cain, B.D., Sielaff, H., Wintermann, F., Engelbrecht, S., Junge, W., Two rotary motors in F-ATP synthase are elastically coupled by a flexible rotor and a stiff stator stalk (2011) Proc. Natl. Acad. Sci. U. S. A., 108, pp. 3924-3929
Stewart, A.G., Lee, L.K., Donohoe, M., Chaston, J.J., Stock, D., The dynamic stator stalk of rotary ATPases (2012) Nat. Commun., 3, p. 687