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J. Biol. Chem., Vol. 283, Issue 37, 25200-25208, September 12, 2008

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The Three Mitochondrial Encoded CcmF Proteins Form a Complex That Interacts with CCMH and c-Type Apocytochromes in Arabidopsis^*

From the Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du général Zimmer, 67084 Strasbourg, France

Three reading frames called ccmF_N1, ccmF_N2, and ccmF_c are found in the mitochondrial genome of Arabidopsis. These sequences are similar to regions of the bacterial gene ccmF involved in cytochrome c maturation. ccmF genes are always absent from animal and fungi genomes but are found in mitochondrial genomes of land plant and several evolutionary distant eukaryotes. In Arabidopsis, ccmF_N2 despite the absence of a classical initiation codon is not a pseudo gene. The 3 ccmF genes of Arabidopsis are expressed at the protein level. Their products are integral proteins of the mitochondrial inner membrane with in total 11 to 13 predicted transmembrane helices. The conserved WWD domain of CcmF_N2 is localized in the inter membrane space. The 3 CcmF proteins are all detected in a high molecular mass complex of 500 kDa by Blue Native PAGE. Direct interaction between CcmF_N2 and both CcmF_N1 and CcmF_C is shown with the yeast two-hybrid split ubiquitin system, but no interaction is observed between CcmF_N1 and CcmF_C. Similarly, interaction is detected between CcmF_N2 and apocytochrome c but also with apocytochrome c₁. Finally, CcmF_N1 and CcmF_N2 both interact with CCMH previously shown to interact as well with cytochrome c. This strengthens the hypothesis that CcmF and CCMH make a complex that performs the assembly of heme with c-type apocytochromes in plant mitochondria.

Received for publication, April 4, 2008 , and in revised form, July 18, 2008.

^* This work was supported in part by the "Centre National de la Recherche Scientifique." The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. S1.

¹ Supported by a Centre Franco-Indien pour la promotion de la Recherche Avancée (CEFIPRA) fellowship. Present address: Avesthagen Limited, Whitefield Road, Bangalore 560066, India.

² Supported by a fellowship of the "Ministère de l'enseignement supérieur et de la recherche."

³ Supported by a European Union Marie Curie reintegration grant and by a "ANR Jeune Chercheur" research grant. To whom correspondence should be addressed. Tel.: 33-0-3-88-41-72-38; Fax: 33-0-3-88-61-44-42; E-mail: Philippe.Giege{at}ibmp-ulp.u-strasbg.fr.

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