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First published online August 29, 2005
doi: 10.1242/10.1242/jcs.02526

Journal of Cell Science 118, 3985-3995 (2005)
Published by The Company of Biologists 2005
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Research Article

Import of rat mitochondrial citrate carrier (CIC) at increasing salt concentrations promotes presequence binding to import receptor Tom20 and inhibits membrane translocation

Vincenzo Zara1,*, Alessandra Ferramosca1, Panagiotis Papatheodorou2, Ferdinando Palmieri3 and Joachim Rassow2

1 Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università di Lecce, I-73100 Lecce, Italy
2 Institut für Mikrobiologie, Universität Hohenheim, 70593 Hohenheim, Germany
3 Dipartimento Farmaco-Biologico, Università di Bari, I-70125 Bari, Italy

* Author for correspondence (e-mail: vincenzo.zara{at}unile.it)

Accepted 7 June 2005

Mitochondria contain a family of related carrier proteins that mediate transport of metabolites across the mitochondrial inner membrane. All members of this family are synthesized in the cytosol. We characterized the interactions of newly synthesized rat citrate carrier (CIC) precursor protein (pCIC) with the components of the mitochondrial protein import machinery. pCIC contains both a positively charged presequence of 13 amino acids and internal targeting sequences. We found that the pCIC presequence does not interfere with the import pathway and merely acts as an internal chaperone in the cytosol. Under conditions of increased ionic strength, the pCIC presequence binds to the import receptor Tom20 and accumulates at the mitochondrial surface, thereby delaying pCIC translocation across the mitochondrial outer membrane. Similarly, the presequence of the bovine phosphate carrier (PiC) precursor protein (pPiC) is arrested at the mitochondrial surface when salt concentrations are elevated. We conclude that presequences can only act as mediators of mitochondrial protein import if they allow rapid release from import receptor sites. Release from receptors sites may be rate-limiting in translocation.

Key words: Citrate carrier, Mitochondria, Membrane translocation


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