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
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
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
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
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
—, 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
—, 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
—, Hofmann S, Neupert W, Brunner M (2000) Protein translocation into mitochondria: the role of TIM complexes. Trends Cell Biol 10: 25–31
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
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
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
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
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
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
—, 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
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
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
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
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
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
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
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
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
—, 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
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
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
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
—, 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
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
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
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
—, 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
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
—, 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
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
—, 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
—, 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
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
—, 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
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
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
—, 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
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
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
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
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
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
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
—, 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
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
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
Nelson DR, Felix CM, Swanson JM (1998) Highly conserved chargepair networks in the mitochondrial carrier family. J Mol Biol 277: 285–308
Neupert W (1997) Protein import into mitochondria. Annu Rev Biochem 66: 863–917
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
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
Pfanner N (1998) Mitochondrial import: crossing the aqueous intermembrane space. Curr Biol 8: R262–265
—, Neupert W (1987) Distinct steps in the import of the ADP/ATP carrier into mitochondria. J Biol Chem 262: 7528–7536
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
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
Ryan KR, Jensen RE (1995) Protein translocation across mitochondrial membranes: what a long, strange trip it is. Cell 83: 517–519
—, 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
—, 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
—, Muller H, Pfanner N (1999) Functional staging of ADP/ATP carrier translocation across the outer mitochondrial membrane. J Biol Chem 274: 20619–20627
Schatz G (1996) The protein import system of mitochondria. J Biol Chem 271: 31763–31766
—, Dobberstein B (1996) Common principles of protein translocation across membranes. Science 271: 1519–1526
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
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
—, 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
Simon SM, Peskin CS, Oster GF (1992) What drives the translocation of proteins? Proc Natl Acad Sei USA 89: 3770–3774
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
—, 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
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
Söllner T, Griffiths G, Pfaller R, Pfanner N, Neupert W (1989) MOM19, an import receptor for mitochondrial precursor proteins. Cell 59: 1061–1070
—, Pfaller R, Griffiths G, Pfanner N, Neupert W (1990) A mitochondrial import receptor for the ATP/ADP carrier. Cell 62: 107–115
Ungermann C, Neupert W, Cyr DM (1994) The role of Hsp70 in conferring unidirectionality on protein translocation into mitochondria. Science 266: 1250–1253
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
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
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
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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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DOI: https://doi.org/10.1007/BF01280499