CGM - DepartmentRegulation and Compartmentalization of Cellular Functions - G. Dujardin's team
Import of mitochondrial proteins
Principal Investigator: B. GUIARD
Last update: 10-Jan-2012
Theme
Ninety nine percent of the 1,000 – 1,500 proteins present in the mitochondria are synthesised in the cytoplasm and imported in the organelle via complicated machineries, such as the complexes TOM and the TIM of the external and the inner mitochondrial membrane, respectively. The majority of the respiratory complexes subunits follow that way.
Schematic model of the mitochondrial import machineries in yeast
S. cerevisiae (Pfanner et al.)
Two new import pathways, called SAM and MIA, have been discovered more recently. We are currently studying the different actors involved in these import machineries, and their molecular mechanisms.
The SAM complex, formed by three proteins, Sam50, Sam35 and Sam37, is involved in the translocation of Beta-barrel proteins, such as the porin and Tom40, in the inner mitochondrial membrane. All the actors of that pathway are likely to have been now identified but their precise mechanisms need to be elucidated. The MIA pathway, with the proteins Mia40 and Erv1, is mainly involved in the import of small cystein-rich proteins in the intermembrane space (IMS). The import and folding of these small proteins are performed through a redox system that involves a transient relay of disulphur brigdes. This is a new concept, an exchange of disulphur bridges leading to the vectorial translocation of proteins through a membrane. The different actors of the MIA pathway and their mode of action have to be discovered.
Most of the proteins involved in mitochondrial import are part of multimeric complexes called translocases. These are dynamic machines. When reacting with the proteins to be imported, they undergo important rearrangements and can interact transiently with each other. We have observed that IMS chaperons cooperate with the translocases TIM and TOM. Other complexes participate in these interactions, such as the respiratory chain complexes and the MDM complex that plays an key role in the control of the mitochondrial morphology, the biogenesis and assembly of the TOM complex and the synthesis of lipids.
We are performing these studies in collaboration with K. Pfanner’s and P. Rehling’s laboratories.
Collaborations
- Nikolaus Pfanner, Albert-Ludwigs-Universität Freiburg, Germany. Study of import pathways in the different compartments of the mitochondria.
- Peter Rehling, University de Göttingen, Germany. Study of the functional coupling of the TOM-TIM translocases.
Publications since 1999
- Gebert, N., ...., Guiard, B., ... van der Laan, M., Pfanner, N., Wiedemann, N. (2011) Dual Function of Sdh3 in the Respiratory Chain and TIM22 Protein Translocase of the Mitochondrial Inner Membrane. Mol Cell, 44 (5) 811-8.
- Schulz, C., Lytovchenko, O., Melin, J., Chacinska, A., Guiard, B., Neumann, P., Ficner, R., Jahn, O., Schmidt, B., Rehling, P. (2011) Tim50's presequence receptor domain is essential for signal driven transport across the TIM23 complex. J Cell Biol, 195 (4) 643-56.
- Stroud, DA., Oeljeklaus, S., Wiese, S., Bohnert, M., Lewandrowski, U., Sickmann, A., Guiard, B., van der Laan, M., Warscheid, B., Wiedemann, N. (2011) Composition and topology of the endoplasmic reticulum-mitochondria encounter structure. J Mol Biol, 413 (4) 743-50.
- Becker, T., Wenz, LS., Krüger, V., Lehmann, W., Müller, JM., Goroncy, L., Zufall, N., Lithgow, T., Guiard, B., Chacinska, A., Wagner, R., Meisinger, C., Pfanner, N. (2011) The mitochondrial import protein Mim1 promotes biogenesis of multispanning outer membrane proteins. J Cell Biol, 194 (3 387-95.
- Stroud, DA., Becker, T., Qiu, J., Stojanovski, D., Pfannschmidt, S., Wirth, C., Hunte, C., Guiard, B., Meisinger, C., Pfanner, N., Wiedemann, N. (2001)Biogenesis of mitochondrial {beta}-barrel proteins: the POTRA domain is involved in precursor release from the SAM complex. Mol Biol Cell, 22 (16) 2823-2833.
- Schmidt, O., Harbauer, AB., Rao, S., Eyrich, B., Zahedi, RP., Stojanovski., D, Schönfisch, B., Guiard, B., Sickmann, A., Pfanner, N., Meisinger, C. (2011) Regulation of mitochondrial protein import by cytosolic kinases. Cell, 144 (2) 227-39.
- Corvest, V., Murrey, DA., Bernard, DG., Knaff, DB., Guiard, B., Hamel, PP. (2010) C-type CytochromeAssembly in Saccharomyces cerevisiae: A Key Residue for Apocytochrome C1/Lyase Interaction. Genetics, 186 (2) 561-71.
- Becker, T., Guiard, B., Thornton, N., Zufall, N., Stroud, DA., Wiedemann, N., Pfanner, N. (2010) Assembly of the Mitochondrial Protein Import Channel: Role of Tom5 in Two-Stage Interaction of Tom40 with the SAM Complex. Mol Biol Cell, 21 (19) 3106-13.
- Bohnert, M., Rehling, P., Guiard, B., Herrmann, JM., Pfanner, N., van der Laan, M. (2010) Cooperation of stop-transfer and conservative sorting mechanisms in mitochondrial protein transport.Curr Biol, 20 (13) 1227-32.
- Chacinska, A., van der Laan, M., Mehnert, C.-S., Guiard, B., Mick, D.-U., Hutu, D.-P., Truscott, K., Wiedemann, N., Meisinger, C., Pfanner, N. and Rehling, P. (2010) Distinct forms of mitochondrial TOM-TIM supercomplexes define signal-dependent states of preprotein sorting. Mol Cell Biol, 30 (1) 307-18.
- Thornton, N., Stroud, D.-A., Milenkovic, D., Guiard, B., Pfanner, N. and Becker, T. (2010) Two Modular Forms of the Mitochondrial Sorting and Assembly Machinery Are Involved in Biogenesis of alpha-Helical Outer Membrane Proteins. J Mol Biol, 396 (3) 540-549.
- Bourges, I., Mucchielli, M.-H., Herbert, C.-J., Guiard, B., Dujardin, G. and Meunier, B. (2009) Multiple Defects in the Respiratory Chain Lead to the Repression of Genes Encoding Components of the Respiratory Chain and TCA Cycle Enzymes. J Mol Biol, 23 (1) 23-42.
- Baker, M. J., Webb, C. T., Stroud, D. A., Palmer, C. S., Frazier, A. E., Guiard, B., Chacinska, A., Gulbis, J. M. and Ryan, M. T. (2009) Structural and functional requirements for activity of the Tim9-Tim10 complex in mitochondrial protein import. Mol Biol Cell, 20 (3) 769-79.
- Vernis, L., Facca, C., Delagoutte, E., Soler, N., Chanet, R., Guiard, B., Faye, G. and Baldacci, G. (2009) A newly identified essential complex, Dre2-Tah18, controls mitochondria integrity and cell death after oxidative stress in yeast. PLoS ONE, 4 (2) e4376.
- Kutik, S., Rissler, M., Guan, X. L., Guiard, B., Shui, G., Gebert, N., Heacock, P. N., Rehling, P., Dowhan, W., Wenk, M. R., Pfanner, N. and Wiedemann, N. (2008) The translocator maintenance protein Tam41 is required for mitochondrial cardiolipin biosynthesis. J Cell Biol, 183 (7) 1213-21.
- Stojanovski, D., Milenkovic, D., Müller, J.-M., Gabriel, K., Schulze-Specking, A., Baker, M.-J., Ryan, M.-T., Guiard, B., Pfanner, N. and Chacinska, A. (2008) Mitochondrial protein import: precursor oxidation in a ternary complex with disulfide carrier and sulfhydryl oxidase. J Cell Biol, 183 (2) 195-202.
- Bao, W.-G., Guiard, B., Fang, Z.-A., Donnini, C., Gervais, M., Passos, F.-M., Ferrero, I., Fukuhara, H. and Bolotin-Fukuhara, M. (2008) Oxygen-dependent transcriptional regulator Hap1p limits glucose uptake by repressing the expression of the major glucose transporter gene RAG1 in Kluyveromyces lactis. Eukaryot Cell, 7 (11) 1895-905.
- Müller, J.-M., Milenkovic, D., Guiard, B., Pfanner, N. and Chacinska, A. (2008) Precursor Oxidation by Mia40 and Erv1 Promotes Vectorial Transport of Proteins into the Mitochondrial Intermembrane Space. Mol Biol Cell, 19 (1) 226-36.
- Chacinska, A., Guiard, B., Müller, J.-M., Schulze-Specking, A., Gabriel, K., Kutik, S. and Pfanner, N. (2008) Mitochondrial biogenesis: Switching the sorting pathway of the intermembrane space receptor Mia40. J Biol Chem, 283 (44) 29723-9.
- Wagner, K., Gebert, N., Guiard, B., Brandner, K., Truscott, K.-N., Wiedemann, N., Pfanner, N. and Rehling, P. (2008) The Assembly Pathway of the Mitochondrial Carrier Translocase Involves four Preprotein Translocases. Mol Cell Biol, 28 (13) 4251-60.
- Hutu, D.-P., Guiard, B., Chacinska, A., Becker, D., Pfanner, N., Rehling, P. and van der Laan, M. (2008) Mitochondrial Protein Import Motor: Differential Role of Tim44 in the Recruitment of Pam17 and J-complex to the Presequence Translocase. Mol Biol Cell, 19 (6) 2642-9.
- Gebert, N., Chacinska, A., Wagner, K., Guiard, B., Koehler, C.-M., Rehling, P., Pfanner, N. and Wiedemann, N. (2008) Assembly of the three small Tim proteins precedes docking to the mitochondrial carrier translocase. EMBO Reports, 9 (6) 548-54.
- Kutik, S., Stojanovski, D., Becker, L., Becker, T., Meinecke, M., Krüger, V., Prinz, C., Meisinger, C., Guiard, B., Wagner, R., Pfanner, N. and Wiedemann, N. (2008) Dissecting membrane insertion of mitochondrial beta-barrel proteins. Cell, 132 (6) 1011-24.
- Becker, T., Pfannschmidt, S., Guiard, B., Stojanovski, D., Milenkovic, D., Kutik, S., Pfanner, N., Meisinger, C. and Wiedemann, N. (2008) Biogenesis of the mitochondrial TOM complex: Mim1 promotes insertion and assembly of signal-anchored receptors. J Biol Chem, 283 (1) 120-127.
- Stojanovski, D., Müller, J.-M., Milenkovic, D., Guiard, B., Pfanner, N. and Chacinska, A. (2008) The MIA system for protein import into the mitochondrial intermembrane space. Biochim Biophys Acta, 1783 (4) 610-7.
- Stojanovski, D., Guiard, B., Kozjak-Pavlovic, V., Pfanner, N. and Meisinger, C. (2007) Alternative function for the mitochondrial SAM complex in biogenesis of {alpha}-helical TOM proteins. J Cell Biol, 179 (81) 881-93.
- Müller, J.-M., Milenkovic, D., Guiard, B., Pfanner, N. and Chacinska, A. (2007) Precursor Oxidation by Mia40 and Erv1 Promotes Vectorial Transport of Proteins into the Mitochondrial Intermembrane Space. Mol Biol Cell, 2007 Oct 31 [Epub ahead of print]
- Kutik, S., Guiard, B., Meyer, H.-E., Wiedemann, N. and Pfanner, N. (2007) Cooperation of translocase complexes in mitochondrial protein import. J Cell Biol, 179 (4) 585-91.
- van der Laan, M., Meinecke, M., Dudek, J., Hutu, D.-P., Lind, M., Perschil, I., Guiard, B., Wagner, R., Pfanner, N. and Rehling, P. (2007) Motor-free mitochondrial presequence translocase drives membrane integration of preproteins. Nat Cell Biol,9 (20) 1152-1159.
- Krayl, M., Lim, J.-H., Martin, F., Guiard, B. and Voos, W. (2007) A cooperative action of the ATP-dependent import motor complex and the inner membrane potential drives mitochondrial preprotein import. Mol Cell Biol, 27 (2) 411-25.
- Gabriel, K., Milenkovic, D., Chacinska, A., Muller, J., Guiard, B., Pfanner, N. and Meisinger, C. (2007) Novel Mitochondrial Intermembrane Space Proteins as Substrates of the MIA Import Pathway. J Mol Biol, 365 (3) 612-20.
- Milenkovic, D., Gabriel, K., Guiard, B., Schulze-Specking, A., Pfanner, N. and Chacinska, A. (2007) Biogenesis of the essential Tim9-Tim10 chaperone complex of mitochondria: site-specific recognition of cysteine residues by the intermembrane space receptor Mia40. J Biol Chem, 282 (31) 22472-80.
- Meisinger, C., Pfannschmidt, S., Rissler, M., Milenkovic, D., Becker, T., Stojanovski, D., Youngman, M. J., Jensen, R.-E., Chacinska, A., Guiard, B., Pfanner, N. and Wiedemann, N. (2007) The morphology proteins Mdm12/Mmm1 function in the major beta-barrel assembly pathway of mitochondria. EMBO J, 26 (9) 2229-39.
- Sanjuan Szklarz, L., Kozjak-Pavlovic, V., Vogtle, F., Chacinska, A., Milenkovic, D., Vogel, S., Durr, M., Westermann, B., Guiard, B., Martinou, J., Borner, C., Pfanner, N. and Meisinger, C. (2007) Preprotein Transport Machineries of Yeast Mitochondrial Outer Membrane Are not Required for Bax-induced Release of Intermembrane Space Proteins. J Mol Biol, 368 (1) 44-54.
- Albrecht, R., Rehling, P., Chacinska, A., Brix, J., Cadamuro, S. A., Volkmer, R., Guiard, B., Pfanner, N. and Zeth, K. (2006) The Tim21 binding domain connects the preprotein translocases of both mitochondrial membranes. EMBO Reports, 7 (12) 1233-8.
- Bernard, D. (2006) Biogenèse des cytochromes de type c dans la mitochondrie chez la levure Saccharomyces cerevisiae. Thèse, Paris Sud 11 Orsay.
- Frazier, A. E., Taylor, R. D., Mick, D. U., Warscheid, B., Stoepel, N., Meyer, H. E., Ryan, M. T., Guiard, B. and Rehling, P. (2006) Mdm38 interacts with ribosomes and is a component of the mitochondrial protein export machinery. J Cell Biol, 172 (4) 553-64.
- Gabriel, K., Milenkovic, D., Chacinska, A., Muller, J., Guiard, B., Pfanner, N. and Meisinger, C. (2006) Novel Mitochondrial Intermembrane Space Proteins as Substrates of the MIA Import Pathway. J Mol Biol, 2006 Oct 14; [Epub ahead of print]
- Meinecke, M., Wagner, R., Kovermann, P., Guiard, B., Mick, D. U., Hutu, D. P., Voos, W., Truscott, K. N., Chacinska, A., Pfanner, N. and Rehling, P. (2006) Tim50 Maintains the Permeability Barrier of the Mitochondrial Inner Membrane. Science, 312 (5779) 1523 -1526.
- van der Laan, M., Wiedemann, N., Mick, D. U., Guiard, B., Rehling, P. and Pfanner, N. (2006) A role for Tim21 in membrane-potential-dependent preprotein sorting in mitochondria. Curr Biol, 16 (22) 2271-6.
- Zahedi, R. P., Sickmann, A., Boehm, A. M., Winkler, C., Zufall, N., Schonfisch, B., Guiard, B., Pfanner, N. and Meisinger, C. (2006) Proteomic analysis of the yeast mitochondrial outer membrane reveals accumulation of a subclass of preproteins. Mol Biol Cell, 17 1436-50.
- Wiedemann, N., Urzica, E., Guiard, B., Muller, H., Lohaus, C., Meyer, HE., Ryan, MT., Meisinger, C., Muhlenhoff, U., Lill, R., Pfanner, N. (2006) Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. EMBO J. 25(1):184-95
- von Janowsky, B., Knapp, K., Major, T., Krayl, M., Guiard, B., and Voos, W. (2005). Structural properties of substrate proteins determine their proteolysis by the mitochondrial AAA+ protease Pim1. Biol Chem, 386, 12, 1307-17. .
- Rissler, M., Wiedemann, N., Pfannschmidt, S., Guiard, B., Pfanner, N., and Chacinska, A. (2005). The essential mitochondrial protein Erv1 cooperates with Mia40 in biogenesis of intermembrane space proteins. J Mol Biol, 353, 3 485-92
- Bernard, D., Quevillon-Cheruel, S., Merchant, S., Guiard, B., and Hamel, P. (2005). Cyc2p, a membrane-bound flavoprotein involved in the maturation of mitochondrial c-type cytochromes. J Biol Chem 280,48,39852-9
- Sanjuan Szklarz, L., Guiard, B., Rissler, M., Wiedemann, N., Kozjak, V., van der Laan, M., Lohaus, C., Marcus, K., Meyer, H., Chacinska, A., Pfanner, N., and Meisinger, C. (2005). Inactivation of the mitochondrial heat shock protein zim17 leads to aggregation of matrix hsp70s followed by pleiotropic effects on morphology and protein biogenesis. J Mol Biol 351, 206-218.
- Seoane, S., Guiard, B., Rodriguez-Torres, A., and Freire-Picos, M. (2005). Effects of splitting alternative KlCYC1 3'-UTR regions on processing: Metabolic consequences and biotechnological applications. J Biotechnol 118, 149-156.
- Sybirna, K., Guiard, B., Li, Y., Bao, W., Bolotin-Fukuhara, M., and Delahodde, A. (2005). A new Hansenula polymorpha HAP4 homologue which contains only the N-terminal conserved domain of the protein is fully functional in Saccharomyces cerevisiae. Curr Genet 47, 172-181.
- van der Laan, M., Chacinska, A., Lind, M., Perschi, I., Sickmann, A., Meyer, H., Guiard, B., Meisinger, C., Pfanner, N., and Rehling, P. (2005). Pam17 is required for architecture and protein import activity of the presequence translocase-associated motor of mitochondria Pam17 is required for architecture and protein import activity of the mitochondrial Hsp70 motor. Mol Cell Biol 25, 7449-7458.
- Chacinska, A., Lind, M., Frazier, A., Dudek, J., Meisinger, C., Geissler, A., A., S., Meyer, H., Truscott, K., Guiard, B., Pfanner, N., and Rehling, P. (2005). Mitochondrial presequence translocase: switching between TOM tethering and motor recruitment involves Tim21 and Tim17. Cell 120, 817-829.
- Frazier, A., Dudek, J., Guiard, B., Voos, W., Li, Y., Lind, M., Meisinger, C., Geissler, A., Sickmann, A., Meyer, H., Bilanchone, V., Cumsky, M., Truscott, K., Pfanner, N., and Rehling, P. (2004). Pam16 has an essential role in the mitochondrial protein import motor. Nat Struct Mol Biol 11, 226-233
- Li, Y., Dudek, J., Guiard, B., Pfanner, N., Rehling, P., and Voos, W. (2004). The presequence translocase-associated protein import motor of mitochondria: Pam16 functions in an antagonistic manner to Pam18. J Biol Chem, 279, 36, 38047-54
- Meisinger, C., Rissler, M., Chacinska, A., Szklarz, L., Milenkovic, D., Kozjak, V., Schonfisch, B., Lohaus, C., Meyer, H., Yaffe, M., Guiard, B., Wiedemann, N., and Pfanner, N. (2004). The mitochondrial morphology protein Mdm10 functions in assembly of the preprotein translocase of the outer membrane. Dev Cell 7, 61-71
- Milenkovic, D., Kozjak, V., Wiedemann, N., Lohaus, C., Meyer, H., Guiard, B., Pfanner, N., and Meisinger, C. (2004). Sam35 of the mitochondrial protein sorting and assembly machinery is a peripheral outer membrane protein essential for cell viability. J Biol Chem 279, 22781-22785
- Wiedemann, N., Truscott, K., Pfannschmidt, S., Guiard, B., Meisinger, C., and Pfanner, N. (2004). Biogenesis of the protein import channel Tom40 of the mitochondrial outer membrane: intermembrane space components are involved in an early stage of the assembly pathway. J Biol Chem 279, 18188-18194
- Buschlen, S., Amillet, J.-M., Guiard, B., Fournier, A., Marcireau, C., and Bolotin-Fukuhara, M. (2003). The S. cerevisiae HAP complex, a key regulator of mitochondrial function, coordinates nuclear and mitochondrial gene expression. Functional and Comparative Genomics 4, 37-46
- Chacinska, A., Rehling, P., Guiard, B., Frazier, A., Schulze-Specking, A., Pfanner, N., Voos, W., and Meisinger, C. (2003). Mitochondrial translocation contact sites: separation of dynamic and stabilizing elements in formation of a TOM-TIM-preprotein supercomplex. EMBO J 22, 5370-5381
- Frazier, A., Chacinska, A., Truscott, K., Guiard, B., Pfanner, N., and Rehling, P. (2003). Mitochondria use different mechanisms for transport of multispanning membrane proteins through the intermembrane space. Mol Cell Biol 23, 7818-7828
- Kozjak, V., Wiedemann, N., Milenkovic, D., Lohaus, C., Meyer, H., Guiard, B., Meisinger, C., and Pfanner, N. (2003). An essential role of sam50 in the protein sorting and assembly machinery of the mitochondrial outer membrane. J Biol Chem 278, 48520-48523
- Sickmann, A., Reinders, J., Wagner, Y., Joppich, C., Zahedi, R., Meyer, H., Schonfisch, B., Perschil, I., Chacinska, A., Guiard, B., Rehling, P., Pfanner, N., and Meisinger, C. (2003). The proteome of Saccharomyces cerevisiae mitochondria. Proc Natl Acad Sci U S A 100, 13207-13212
- Truscott, K., Voos, W., Frazier, A., Lind, M., Li, Y., Dudek, J., Muller, H., Sickmann, A., Meyer, H., Meisinger, C., Guiard, B., Rehling, P., and Pfanner, N. (2003). A J-protein is an essential subunit of the presequence translocase-associated protein import motor of mitochondria. J Cell Biol 163, 707-713
- Kovermann P, Truscott KN, Guiard B, Rehling P, Sepuri NB, Muller H, Jensen RE, Wagner R, Pfanner N. Tim22, the essential core of the mitochondrial protein insertion complex, forms a voltage-activated and signal-gated channel. Mol Cell. 2002 Feb;9(2):363-73.
- Lodi T, Fontanesi F, Guiard B. Co-ordinate regulation of lactate metabolism genes in yeast: the role of the lactate permease gene JEN1. Mol Genet Genomics. 2002 Jan;266(5):838-47
- Lim JH, Martin F, Guiard B, Pfanner N, Voos W. Related Articles The mitochondrial Hsp70-dependent import system actively unfolds preproteins and shortens the lag phase of translocation. EMBO J. 2001 Mar 1;20(5):941-50.
- Ramil E, Agrimonti C, Shechter E, Gervais M, Guiard B. Regulation of the CYB2 gene expression: transcriptional co-ordination by the hap1p, Hap2/3/4/5p and adr1p transcription factors. Mol Microbiol. 2000 Sep;37(5):1116-32.
- Leonhard K, Guiard B, Pellecchia G, Tzagoloff A, Neupert W, Langer T. Membrane protein degradation by AAA proteases in mitochondria: extraction of substrates from either membrane surface. Mol Cell. 2000 Apr;5(4):629-38.
- Donzeau M, Kaldi K, Adam A, Paschen S, Wanner G, Guiard B, Bauer MF, Neupert W, Brunner M. Tim23 links the inner and outer mitochondrial membranes. Cell. 2000 May 12;101(4):401-12.
- Alberti A, Guiard B, Ferrero I. Regulation of the Saccharomyces cerevisiae DLD1 gene encoding the mitochondrial protein D-lactate ferricytochrome c by HAP1 and HAP2/3/4/5. Mol Gen Genet. 1999 Dec;262(4-5):623-32.
- van Wilpe S, Ryan MT, Hill K, Maarse AC, Meisinger C, Brix J, Dekker PJ, Moczko M, Wagner R, Meijer M, Guiard B, Honlinger A, Pfanner N. Tom22 is a multifunctional organizer of the mitochondrial preprotein translocase. Nature. 1999 Sep 30;401(6752):485-9.
- Diekert K, Kispal G, Guiard B, Lill R. An internal targeting signal directing proteins into the mitochondrial intermembrane space. Proc Natl Acad Sci U S A. 1999 Oct 12;96(21):11752-7.
- Rojo EE, Guiard B, Neupert W, Stuart RA. N-terminal tail export from the mitochondrial matrix. Adherence to the prokaryotic "positive-inside" rule of membrane protein topology. J Biol Chem. 1999 Jul 9;274(28): 19617-22.
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