Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1998 Mar 16;17(6):1569–1576. doi: 10.1093/emboj/17.6.1569

Biogenesis of Tim23 and Tim17, integral components of the TIM machinery for matrix-targeted preproteins.

K Káldi 1, M F Bauer 1, C Sirrenberg 1, W Neupert 1, M Brunner 1
PMCID: PMC1170504  PMID: 9501078

Abstract

We analysed the import pathway of Tim23 and of Tim17, components of the mitochondrial import machinery for matrix-targeted preproteins. Tim23 contains two independent import signals. One is located within the first 62 amino acid residues of the hydrophilic domain that, in the assembled protein, is exposed to the intermembrane space. This signal mediates translocation of Tim23 across the outer membrane independently of the membrane potential, DeltaPsi. A second import signal is located in the C-terminal membrane-integrated portion of Tim23. It mediates translocation across the outer membrane and insertion into the inner membrane in a strictly DeltaPsi-dependent fashion. Structurally, Tim17 is related to Tim23 but lacks a hydrophilic domain. It contains an import signal in the C-terminal half and its import requires DeltaPsi. The DeltaPsi-dependent import signals of Tim23 and Tim17 are located at corresponding sites in these two homologous proteins. They exhibit features reminiscent of the positively charged N-terminal presequences of matrix-targeted precursors. Import of Tim23 and its insertion into the inner membrane requires Tim22 but not functional Tim23. Thus, biogenesis of the Tim23.17 complex depends on the Tim22 complex, which is the translocase identified as mediating the import of carrier proteins.

Full Text

The Full Text of this article is available as a PDF (316.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bauer M. F., Sirrenberg C., Neupert W., Brunner M. Role of Tim23 as voltage sensor and presequence receptor in protein import into mitochondria. Cell. 1996 Oct 4;87(1):33–41. doi: 10.1016/s0092-8674(00)81320-3. [DOI] [PubMed] [Google Scholar]
  2. Berthold J., Bauer M. F., Schneider H. C., Klaus C., Dietmeier K., Neupert W., Brunner M. The MIM complex mediates preprotein translocation across the mitochondrial inner membrane and couples it to the mt-Hsp70/ATP driving system. Cell. 1995 Jun 30;81(7):1085–1093. doi: 10.1016/s0092-8674(05)80013-3. [DOI] [PubMed] [Google Scholar]
  3. Bolliger L., Junne T., Schatz G., Lithgow T. Acidic receptor domains on both sides of the outer membrane mediate translocation of precursor proteins into yeast mitochondria. EMBO J. 1995 Dec 15;14(24):6318–6326. doi: 10.1002/j.1460-2075.1995.tb00322.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Court D. A., Nargang F. E., Steiner H., Hodges R. S., Neupert W., Lill R. Role of the intermembrane-space domain of the preprotein receptor Tom22 in protein import into mitochondria. Mol Cell Biol. 1996 Aug;16(8):4035–4042. doi: 10.1128/mcb.16.8.4035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dekker P. J., Keil P., Rassow J., Maarse A. C., Pfanner N., Meijer M. Identification of MIM23, a putative component of the protein import machinery of the mitochondrial inner membrane. FEBS Lett. 1993 Sep 6;330(1):66–70. doi: 10.1016/0014-5793(93)80921-g. [DOI] [PubMed] [Google Scholar]
  6. Emtage J. L., Jensen R. E. MAS6 encodes an essential inner membrane component of the yeast mitochondrial protein import pathway. J Cell Biol. 1993 Sep;122(5):1003–1012. doi: 10.1083/jcb.122.5.1003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fölsch H., Guiard B., Neupert W., Stuart R. A. Internal targeting signal of the BCS1 protein: a novel mechanism of import into mitochondria. EMBO J. 1996 Feb 1;15(3):479–487. [PMC free article] [PubMed] [Google Scholar]
  8. Glick B., Wachter C., Schatz G. Protein import into mitochondria: two systems acting in tandem? Trends Cell Biol. 1991 Oct;1(4):99–103. doi: 10.1016/0962-8924(91)90037-a. [DOI] [PubMed] [Google Scholar]
  9. Hill J., Donald K. A., Griffiths D. E., Donald G. DMSO-enhanced whole cell yeast transformation. Nucleic Acids Res. 1991 Oct 25;19(20):5791–5791. doi: 10.1093/nar/19.20.5791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horst M., Hilfiker-Rothenfluh S., Oppliger W., Schatz G. Dynamic interaction of the protein translocation systems in the inner and outer membranes of yeast mitochondria. EMBO J. 1995 May 15;14(10):2293–2297. doi: 10.1002/j.1460-2075.1995.tb07223.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hovland P., Flick J., Johnston M., Sclafani R. A. Galactose as a gratuitous inducer of GAL gene expression in yeasts growing on glucose. Gene. 1989 Nov 15;83(1):57–64. doi: 10.1016/0378-1119(89)90403-4. [DOI] [PubMed] [Google Scholar]
  12. Klaus C., Guiard B., Neupert W., Brunner M. Determinants in the presequence of cytochrome b2 for import into mitochondria and for proteolytic processing. Eur J Biochem. 1996 Mar 15;236(3):856–861. doi: 10.1111/j.1432-1033.1996.00856.x. [DOI] [PubMed] [Google Scholar]
  13. Koll H., Guiard B., Rassow J., Ostermann J., Horwich A. L., Neupert W., Hartl F. U. Antifolding activity of hsp60 couples protein import into the mitochondrial matrix with export to the intermembrane space. Cell. 1992 Mar 20;68(6):1163–1175. doi: 10.1016/0092-8674(92)90086-r. [DOI] [PubMed] [Google Scholar]
  14. Kronidou N. G., Oppliger W., Bolliger L., Hannavy K., Glick B. S., Schatz G., Horst M. Dynamic interaction between Isp45 and mitochondrial hsp70 in the protein import system of the yeast mitochondrial inner membrane. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12818–12822. doi: 10.1073/pnas.91.26.12818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lill R., Stuart R. A., Drygas M. E., Nargang F. E., Neupert W. Import of cytochrome c heme lyase into mitochondria: a novel pathway into the intermembrane space. EMBO J. 1992 Feb;11(2):449–456. doi: 10.1002/j.1460-2075.1992.tb05074.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lithgow T., Glick B. S., Schatz G. The protein import receptor of mitochondria. Trends Biochem Sci. 1995 Mar;20(3):98–101. doi: 10.1016/s0968-0004(00)88972-0. [DOI] [PubMed] [Google Scholar]
  17. Maarse A. C., Blom J., Keil P., Pfanner N., Meijer M. Identification of the essential yeast protein MIM17, an integral mitochondrial inner membrane protein involved in protein import. FEBS Lett. 1994 Aug 1;349(2):215–221. doi: 10.1016/0014-5793(94)00669-5. [DOI] [PubMed] [Google Scholar]
  18. Mayer A., Nargang F. E., Neupert W., Lill R. MOM22 is a receptor for mitochondrial targeting sequences and cooperates with MOM19. EMBO J. 1995 Sep 1;14(17):4204–4211. doi: 10.1002/j.1460-2075.1995.tb00094.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mayer A., Neupert W., Lill R. Mitochondrial protein import: reversible binding of the presequence at the trans side of the outer membrane drives partial translocation and unfolding. Cell. 1995 Jan 13;80(1):127–137. doi: 10.1016/0092-8674(95)90457-3. [DOI] [PubMed] [Google Scholar]
  20. Neupert W. Protein import into mitochondria. Annu Rev Biochem. 1997;66:863–917. doi: 10.1146/annurev.biochem.66.1.863. [DOI] [PubMed] [Google Scholar]
  21. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  22. Pfanner N., Rassow J., van der Klei I. J., Neupert W. A dynamic model of the mitochondrial protein import machinery. Cell. 1992 Mar 20;68(6):999–1002. doi: 10.1016/0092-8674(92)90069-o. [DOI] [PubMed] [Google Scholar]
  23. Rassow J., Maarse A. C., Krainer E., Kübrich M., Müller H., Meijer M., Craig E. A., Pfanner N. Mitochondrial protein import: biochemical and genetic evidence for interaction of matrix hsp70 and the inner membrane protein MIM44. J Cell Biol. 1994 Dec;127(6 Pt 1):1547–1556. doi: 10.1083/jcb.127.6.1547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ryan K. R., Jensen R. E. Mas6p can be cross-linked to an arrested precursor and interacts with other proteins during mitochondrial protein import. J Biol Chem. 1993 Nov 15;268(32):23743–23746. [PubMed] [Google Scholar]
  25. Ryan K. R., Menold M. M., Garrett S., Jensen R. E. SMS1, a high-copy suppressor of the yeast mas6 mutant, encodes an essential inner membrane protein required for mitochondrial protein import. Mol Biol Cell. 1994 May;5(5):529–538. doi: 10.1091/mbc.5.5.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schneider H. C., Berthold J., Bauer M. F., Dietmeier K., Guiard B., Brunner M., Neupert W. Mitochondrial Hsp70/MIM44 complex facilitates protein import. Nature. 1994 Oct 27;371(6500):768–774. doi: 10.1038/371768a0. [DOI] [PubMed] [Google Scholar]
  27. Schneider H. C., Westermann B., Neupert W., Brunner M. 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. 1996 Nov 1;15(21):5796–5803. [PMC free article] [PubMed] [Google Scholar]
  28. Sirrenberg C., Bauer M. F., Guiard B., Neupert W., Brunner M. Import of carrier proteins into the mitochondrial inner membrane mediated by Tim22. Nature. 1996 Dec 12;384(6609):582–585. doi: 10.1038/384582a0. [DOI] [PubMed] [Google Scholar]
  29. Sirrenberg C., Endres M., Becker K., Bauer M. F., Walther E., Neupert W., Brunner M. Functional cooperation and stoichiometry of protein translocases of the outer and inner membranes of mitochondria. J Biol Chem. 1997 Nov 21;272(47):29963–29966. doi: 10.1074/jbc.272.47.29963. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES