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Published/Copyright: July 27, 2005
Biological Chemistry
From the journal Volume 381 Issue 7

Abstract

Import of DNA from the cytoplasm into the mitochondrial matrix is an obligatory step for an in organello sitedirected mutagenesis or gene therapy approach on mitochondrial DNA diseases. In this context, we have developed an artificial DNA translocation vector that is composed of the mitochondrial signal peptide of the ornithine transcarbamylase (OTC) and a DNA moiety. While this vector is capable of directing attached passenger molecules to the mitochondrial matrix, the recognition of this artificial molecule by the endogenous mitochondrial signal peptide processing machinery as well as the cleavage of the peptide plays a pivotal role in the release of the attached DNA. To study the proteolytic processing of the artificial vector, various signal peptideDNAconjugates were treated with purified mitochondrial intermediate peptidase. When the leader peptide is directly linked to the DNA moiety without an intervening spacer, MIP processing is prevented. Cleavage of the peptide can be restored, however, when the first ten amino acid residues of the mature part of OTC are appended at the carboxyterminal end of the signal peptide. Our results show that artificial peptideDNAconjugates are recognized by the mitochondrial proteolytic machinery, and therefore an interference of the peptide with the DNA function can be excluded.

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Published Online: 2005-07-27
Published in Print: 2000-07-04

Copyright © 2000 by Walter de Gruyter GmbH & Co. KG

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