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Mechanisms of mitochondrial protein import

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Summary

Import of nuclear encoded precursor proteins into and across the mitochondrial inner membrane is mediated by two distinct translocases, the TIM23 complex and the TIM22 complex, which both cooperate with the general preprotein translocase of the outer membrane, TOM complex. The TIM23 complex mediates import of preproteins with a positively charged matrix-targeting signal, while the TIM22 complex mediates the import of a class of integral inner-membrane proteins which do not carry a matrixtargeting signal. The focus of this review is on the structural organization and function of components that act along these import pathways and their conservation between species.

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References

  • Abe Y, Shodai T, Muto T, Mihara K, Torii H, Nishikawa S, Endo T, Kohda D (2000) Structural basis of presequence recognition by the mitochondrial protein import receptor Tom20. Cell 100: 551–560

    Google Scholar 

  • Adam A, Endres M, Sirrenberg C, Lottspeich F, Neupert W, Brunner M (1999) Tim9, a new component of the TIM22.54 translocase in mitochondria. EMBO J 18: 313–319

    Google Scholar 

  • Ahting U, Thun C, Hegerl R, Typke D, Nargang FE, Neupert W, Nussberger S (1999) The TOM core complex: the general protein import pore of the outer membrane of mitochondria. J Cell Biol 147: 959–968

    Google Scholar 

  • Alconada A, Kübrich M, Moczko M, Hönlinger A, Pfanner N (1995) The mitochondrial receptor complex: the small subunit Mom8b/Isp6 supports association of receptors with the general insertion pore and transfer of preproteins. Mol Cell Biol 15: 6196–6205

    Google Scholar 

  • Bauer MF, Sirrenberg C, Neupert W, Brunner M (1996) Role of Tim23 as voltage sensor and presequence receptor in protein import into mitochondria. Cell 87: 33–41

    Google Scholar 

  • —, Gempel K, Reichert AS, Rappold GA, Lichtner P, Gerbitz KD, Neupert W, Brunner M, Hofmann S (1999a) Genetic and structural characterization of the human mitochondrial inner membrane translocase. J Mol Biol 289: 69–82

    Google Scholar 

  • —, Rothbauer U, Mühlenbein N, Smith RJ, Gerbitz K, Neupert W, Brunner M, Hofmann S (1999b) The mitochondrial TIM22 preprotein translocase is highly conserved throughout the eukaryotic kingdom. FEBS Lett 464: 41–47

    Google Scholar 

  • —, Hofmann S, Neupert W, Brunner M (2000) Protein translocation into mitochondria: the role of TIM complexes. Trends Cell Biol 10: 25–31

    Google Scholar 

  • Berthold J, Bauer MF, Schneider HC, Klaus C, Dietmeier K, Neupert W, Brunner M (1995) The MIM complex mediates preprotein translocation across the mitochondrial inner membrane and couples it to the mt-Hsp70/ATP driving system. Cell 81: 1085–1093

    Google Scholar 

  • Blom J, Dekker P, Meijer M (1995) Functional and physical interactions of components of the yeast mitochondrial innermembrane import machinery (MIM). Eur J Biochem 232: 309–314

    Google Scholar 

  • Bomer U, Rassow J, Zufall N, Pfanner N, Meijer M, Maarse AC (1996) The preprotein translocase of the inner mitochondrial membrane: evolutionary conservation of targeting and assembly of Tim17. J Mol Biol 262: 389–395

    Google Scholar 

  • Brix J, Dietmeier K, Pfanner N (1997) Differential recognition of preproteins by the purified cytosolic domains of the mitochondrial import receptors Tom20, Tom22, and Tom70. J Biol Chem 272: 20730–20735

    Google Scholar 

  • Cao W, Douglas MG (1995) Biogenesis of ISP6, a small carboxylterminal anchored protein of the receptor complex of the mitochondrial outer membrane. J Biol Chem 270: 5674–5679

    Google Scholar 

  • Dekker PJ, Keil P, Rassow J, Maarse AC, Pfanner N, Meijer M (1993) Identification of MIM23, a putative component of the protein import machinery of the mitochondrial inner membrane. FEBS Lett 330: 66–70

    Google Scholar 

  • —, Martin F, Maarse AC, Borner U, Muller H, Guiard B, Meijer M, Rassow J, Pfanner N (1997) The Tim core complex defines the number of mitochondrial translocation contact sites and can hold arrested preproteins in the absence of matrix Hsp70-Tim44. EMBO J 16: 5408–5419

    Google Scholar 

  • Dietmeier K, Honlinger A, Bomer U, Dekker PJ, Eckerskorn C, Lottspeich F, Kubrich M, Pfanner N (1997) Tom5 functionally links mitochondrial preprotein receptors to the general import pore. Nature 388: 195–200

    Google Scholar 

  • Donzeau D, Káldi K, Adam A, Paschen S, Wanner G, Guiard B, Bauer MF, Neupert W, Brunner M (2000) Tim23 links the inner and outer mitochondrial membranes. Cell 101: 401–412

    Google Scholar 

  • Emtage JL, Jensen RE (1993) MAS6 encodes an essential inner membrane component of the yeast mitochondrial protein import pathway. J Cell Biol 122: 1003–1012

    Google Scholar 

  • Endres M, Neupert W, Brunner M (1999) Transport of the ADP/ATP carrier of mitochondria from the TOM complex to the TIM22·54 complex. EMBO J (in press)

  • Glick BS (1995) Can Hsp70 proteins act as force-generating motors? Cell 80: 11–14

    Google Scholar 

  • Gratzer S, Lithgow T, Bauer RE, Lamping E, Paltauf F, Kohlwein SD, Haucke V, Junne T, Schatz G, Horst M (1995) Mas37p, a novel receptor subunit for protein import into mitochondria. J Cell Biol 129: 25–34

    Google Scholar 

  • Hachiya N, Mihara K, Suda K, Horst M, Schatz G, Lithgow T (1995) Reconstitution of the initial steps of mitochondrial protein import. Nature 376: 705–709

    Google Scholar 

  • Hill K, Model K, Ryan MT, Dietmeier K, Martin F, Wagner R, Pfanner N (1998) Tom40 forms the hydrophilic channel of the mitochondrial import pore for preproteins. Nature 395: 516–521

    Google Scholar 

  • Hines V, Schatz G (1993) Precursor binding to yeast mitochondria: a general role for the outer membrane protein Mas70p. J Biol Chem 268: 449–454

    Google Scholar 

  • —, Brandt A, Griffiths G, Horstmann H, Brutsch H, Schatz G (1990) Protein import into yeast mitochondria is accelerated by the outer membrane protein MAS70. EMBO J 9: 3191–3200

    Google Scholar 

  • Honlinger A, Bomer U, Alconada A, Eckerskorn C, Lottspeich F, Dietmeier K, Pfanner N (1996) Tom7 modulates the dynamics of the mitochondrial outer membrane translocase and plays a pathway-related role in protein import. EMBO J 15: 2125–2137

    Google Scholar 

  • Ishihara N, Mihara K (1998) Identification of the protein import components of the rat mitochondrial inner membrane, rTIM17, rTIM23, and rTIM44. J Biochem (Tokyo) 123: 722–732

    Google Scholar 

  • Jarosch E, Tuller G, Daum G, Waldherr M, Voskova A, Schweyen RJ (1996) Mrs5p, an essential protein of the mitochondrial intermembrane space, affects protein import into yeast mitochondria. J Biol Chem 271: 17219–17225

    Google Scholar 

  • —, Rodel G, Schweyen RJ (1997) A soluble 12-kDa protein of the mitochondrial intermembrane space, Mrs11p, is essential for mitochondrial biogenesis and viability of yeast cells. Mol Gen Genet 255: 157–165

    Google Scholar 

  • Kaldi K, Bauer MF, Sirrenberg C, Neupert W, Brunner M (1998) Biogenesis of Tim23 and Tim17, integral components of the TIM machinery for matrix-targeted preproteins. EMBO J 17: 1569–1576

    Google Scholar 

  • Kassenbrock CK, Cao W, Douglas MG (1993) Genetic and biochemical characterization of ISP6, a small mitochondrial outer membrane protein associated with the protein translocation complex. EMBO J 12: 3023–3034

    Google Scholar 

  • Kerscher O, Holder J, Srinivasan M, Leung RS, Jensen RE (1997) The Tim54p-Tim22p complex mediates insertion of proteins into the mitochondrial inner membrane. J Cell Biol 139: 1663–1675

    Google Scholar 

  • —, Sepuri NB, Jensen RE (2000) Tim18p is a new component of the Tim54p-Tim22p translocon in the mitochondrial inner membrane. Mol Biol Cell 11: 103–116

    Google Scholar 

  • Kiebler M, Pfaller R, Sollner T, Griffiths G, Horstmann H, Pfanner N, Neupert W (1990) Identification of a mitochondrial receptor complex required for recognition and membrane insertion of precursor proteins. Nature 348: 610–616

    Google Scholar 

  • —, Keil P, Schneider H, van der Klei IJ, Pfanner N, Neupert W (1993) The mitochondrial receptor complex: a central role of MOM22 in mediating preprotein transfer from receptors to the general insertion pore. Cell 74: 483–492

    Google Scholar 

  • Koehler CM, Jarosch E, Tokatlidis K, Schmid K, Schweyen RJ, Schatz G (1998a) Import of mitochondrial carriers mediated by essential proteins of the intermembrane space. Science 279: 369–373

    Google Scholar 

  • —, Merchant S, Oppliger W, Schmid K, Jarosch E, Dolfini L, Junne T, Schatz G, Tokatlidis K (1998b) Tim9p, an essential partner subunit of Tim10p for the import of mitochondrial carrier proteins. EMBO J 17: 6477–6486

    Google Scholar 

  • —, Leuenberger D, Merchant S, Renold A, Junne T, Schatz G (1999) Human deafness dystonia syndrome is a mitochondrial disease. Proc Natl Acad Sei USA 96: 2141–2146

    Google Scholar 

  • Komiya T, Mihara K (1996) Protein import into mammalian mitochondria: characterization of the intermediates along the import pathway of the precursor into the matrix. J Biol Chem 271: 22105–22110

    Google Scholar 

  • —, Rospert S, Schatz G, Mihara K (1997) Binding of mitochondrial precursor proteins to the cytoplasmic domains of the import receptors Tom70 and Tom20 is determined by cytoplasmic chaperones. EMBO J 16: 4267–4275

    Google Scholar 

  • Kronidou NG, Oppliger W, Bolliger L, Hannavy K, Glick BS, Schatz G, Horst M (1994) Dynamic interaction between Isp45 and mitochondrial hsp70 in the protein import system of the yeast mitochondrial inner membrane. Proc Natl Acad Sei USA 91: 12818–12822

    Google Scholar 

  • Kubrich M, Keil P, Rassow J, Dekker PJ, Blom J, Meijer M, Pfanner N (1994) The polytopic mitochondrial inner membrane proteins MIM17 and MIM23 operate at the same preprotein import site. FEBS Lett 349: 222–228

    Google Scholar 

  • —, Rassow J, Voos W, Pfanner N, Honlinger A (1998) The import route of ADP/ATP carrier into mitochondria separates from the general import pathway of cleavable preproteins at the trans side of the outer membrane. J Biol Chem 273: 16374–16381

    Google Scholar 

  • Kunkele KP, Heins S, Dembowski M, Nargang FE, Benz R, Thieffry M, Walz J, Lill R, Nussberger S, Neupert W (1998) The preprotein translocation channel of the outer membrane of mitochondria. Cell 93: 1009–1019

    Google Scholar 

  • Lithgow T, Junne T, Suda K, Gratzer S, Schatz G (1994) The mitochondrial outer membrane protein Mas22p is essential for protein import and viability of yeast. Proc Natl Acad Sei USA 91: 11973–11977

    Google Scholar 

  • Lohret TA, Jensen RE, Kinnally KW (1997) Tim23, a protein import component of the mitochondrial inner membrane, is required for normal activity of the multiple conductance channel, MCC. J Cell Biol 137: 377–386

    Google Scholar 

  • Maarse AC, Blom J, Keil P, Pfanner N, Meijer M (1994) Identification of the essential yeast protein MIM17, an integral mitochondrial inner membrane protein involved in protein import. FEBS Lett 349: 215–221

    Google Scholar 

  • Matouschek A, Azem A, Ratliff K, Glick BS, Schmid K, Schatz G (1997) Active unfolding of precursor proteins during mitochondrial protein import. EMBO J 16: 6727–6736

    Google Scholar 

  • Mayer A, Nargang FE, Neupert W, Lill R (1995a) MOM22 is a receptor for mitochondrial targeting sequences and cooperates with MOM19. EMBO J 14: 4204–4211

    Google Scholar 

  • —, Neupert W, Lill R (1995b) Mitochondrial protein import: reversible binding of the presequence at the trans side of the outer membrane drives partial translocation and unfolding. Cell 80: 127–137

    Google Scholar 

  • Moro F, Sirrenberg C, Schneider H-C, Neupert W, Brunner M (1999) The TIM17·23 preprotein translocase of mitochondria: composition and function in protein transport into the matrix. EMBO J 18: 3667–3675

    Google Scholar 

  • Nakai M, Endo T (1995) Identification of yeast MAS17 encoding the functional counterpart of the mitochondrial receptor complex protein MOM22 ofNeurospora crassa. FEBS Lett 357: 202–206

    Google Scholar 

  • Nelson DR, Felix CM, Swanson JM (1998) Highly conserved chargepair networks in the mitochondrial carrier family. J Mol Biol 277: 285–308

    Google Scholar 

  • Neupert W (1997) Protein import into mitochondria. Annu Rev Biochem 66: 863–917

    Google Scholar 

  • Palmisano A, Zara V, Honlinger A, Vozza A, Dekker PJ, Pfanner N, Palmieri F (1998) Targeting and assembly of the oxoglutarate carrier: general principles for biogenesis of carrier proteins of the mitochondrial inner membrane. Biochem J 333: 151–158

    Google Scholar 

  • Pfaller R, Steger HF, Rassow J, Pfanner N, Neupert W (1988) Import pathways of precursor proteins into mitochondria: multiple receptor sites are followed by a common insertion site. J Cell Biol 107: 2483–2490

    Google Scholar 

  • Pfanner N (1998) Mitochondrial import: crossing the aqueous intermembrane space. Curr Biol 8: R262–265

    Google Scholar 

  • —, Neupert W (1987) Distinct steps in the import of the ADP/ATP carrier into mitochondria. J Biol Chem 262: 7528–7536

    Google Scholar 

  • Ramage L, Junne T, Hahne K, Lithgow T, Schatz G (1993) Functional cooperation of mitochondrial protein import receptors in yeast. EMBO J 12: 4115–4123

    Google Scholar 

  • Rassow J, Maarse AC, Krainer E, Kübrich M, Müller H, Meijer M, Craig EA, Pfanner N (1994) Mitochondrial protein import: biochemical and genetic evidence for interaction of matrix hsp70 and the inner membrane protein MIM44. J Cell Biol 127: 1547–1556

    Google Scholar 

  • Ryan KR, Jensen RE (1995) Protein translocation across mitochondrial membranes: what a long, strange trip it is. Cell 83: 517–519

    Google Scholar 

  • —, Menold MM, Garrett S, Jensen RE (1994) SMS1, a high-copy suppressor of the yeast maso mutant, encodes an essential inner membrane protein required for mitochondrial protein import. Mol Biol Cell 5: 529–538

    Google Scholar 

  • —, Leung RS, Jensen RE (1998) Characterization of the mitochondrial inner membrane translocase complex: the Tim23p hydrophobic domain interacts with Tim17p but not with other Tim23p molecules. Mol Cell Biol 18: 178–187

    Google Scholar 

  • —, Muller H, Pfanner N (1999) Functional staging of ADP/ATP carrier translocation across the outer mitochondrial membrane. J Biol Chem 274: 20619–20627

    Google Scholar 

  • Schatz G (1996) The protein import system of mitochondria. J Biol Chem 271: 31763–31766

    Google Scholar 

  • —, Dobberstein B (1996) Common principles of protein translocation across membranes. Science 271: 1519–1526

    Google Scholar 

  • Schlossmann J, Lill R, Neupert W, Court DA (1996) Tom71, a novel homologue of the mitochondrial preprotein receptor Tom70. J Biol Chem 271: 17890–17895

    Google Scholar 

  • Schneider H-C, Berthold J, Bauer MF, Dietmeier K, Guiard B, Brunner M, Neupert W (1994) Mitochondrial Hsp70/MIM44 complex facilitates protein import. Nature 371: 768–774

    Google Scholar 

  • —, Westermann B., Neupert W, Brunner M (1996) The nucleotide exchange factor MGE exerts a key function in the ATP-dependent cycle of mt-Hsp70-Tim44 interaction driving mitochondrial protein import. EMBO J 15: 5796–5803

    Google Scholar 

  • Simon SM, Peskin CS, Oster GF (1992) What drives the translocation of proteins? Proc Natl Acad Sei USA 89: 3770–3774

    Google Scholar 

  • Sirrenberg C, Bauer MF, Guiard B, Neupert W, Brunner M (1996) Import of carrier proteins into the mitochondrial inner membrane mediated by Tim22. Nature 384: 582–585

    Google Scholar 

  • —, Endres M, Folsch H, Stuart RA, Neupert W, Brunner M (1998) Carrier protein import into mitochondria mediated by the intermembrane proteins Tim1O/Mrs11 and Tim12/Mrs5. Nature 391: 912–915

    Google Scholar 

  • Smagula CS, Douglas MG (1988) ADP/ATP carrier of S. cerevisiae contains a mitochondrial import signal between amino acids 72 and 111. J Cell Biochem 36: 323–328

    Google Scholar 

  • Söllner T, Griffiths G, Pfaller R, Pfanner N, Neupert W (1989) MOM19, an import receptor for mitochondrial precursor proteins. Cell 59: 1061–1070

    Google Scholar 

  • —, Pfaller R, Griffiths G, Pfanner N, Neupert W (1990) A mitochondrial import receptor for the ATP/ADP carrier. Cell 62: 107–115

    Google Scholar 

  • Ungermann C, Neupert W, Cyr DM (1994) The role of Hsp70 in conferring unidirectionality on protein translocation into mitochondria. Science 266: 1250–1253

    Google Scholar 

  • Vestweber D, Brunner J, Baker A, Schatz G (1989) A 42K outermembrane protein is a component of the yeast mitochondrial protein import site. Nature 341: 205–209

    Google Scholar 

  • Voisine C, Craig EA, Zufall N, von Ahsen O, Pfanner N, Voos W (1999) The protein import motor of mitochondria: unfolding and trapping of preproteins are distinct and separable functions of matrix Hsp70. Cell 97: 565–574

    Google Scholar 

  • Voos W, von Ahsen O, Muller H, Guiard B, Rassow J, Pfanner N (1996) Differential requirement for the mitochondrial Hsp70-Tim44 complex in unfolding and translocation of preproteins. EMBO J 15: 2668–2677

    Google Scholar 

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Bauer, M.F., Paschen, S., Neupert, W. et al. Mechanisms of mitochondrial protein import. Protoplasma 213, 1–10 (2000). https://doi.org/10.1007/BF01280499

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