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Originally published In Press as doi:10.1074/jbc.M109993200 on December 14, 2001

J. Biol. Chem., Vol. 277, Issue 8, 6051-6058, February 22, 2002
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The Typically Mitochondrial DNA-encoded ATP6 Subunit of the F1F0-ATPase Is Encoded by a Nuclear Gene in Chlamydomonas reinhardtii*

Soledad FunesDagger §, Edgar Davidson, M. Gonzalo Claros||, Robert van LisDagger **, Xochitl Pérez-MartínezDagger Dagger Dagger , Miriam Vázquez-AcevedoDagger , Michael P. King, and Diego González-HalphenDagger §§

From the Dagger  Departamento de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Apartado Postal 70-243, 04510 México D.F., Mexico, the  Department of Biochemistry and Molecular Pharmacology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, and the || Departamento de Biología Molecular y Bioquímica, Facultad de Ciencias, Universidad de Málaga, Málaga E-29071, Spain

The atp6 gene, encoding the ATP6 subunit of F1F0-ATP synthase, has thus far been found only as an mtDNA-encoded gene. However, atp6 is absent from mtDNAs of some species, including that of Chlamydomonas reinhardtii. Analysis of C. reinhardtii expressed sequence tags revealed three overlapping sequences that encoded a protein with similarity to ATP6 proteins. PCR and 5'- and 3'-RACE were used to obtain the complete cDNA and genomic sequences of C. reinhardtii atp6. The atp6 gene exhibited characteristics of a nucleus-encoded gene: Southern hybridization signals consistent with nuclear localization, the presence of introns, and a codon usage and a polyadenylation signal typical of nuclear genes. The corresponding ATP6 protein was confirmed as a subunit of the mitochondrial F1F0-ATP synthase from C. reinhardtii by N-terminal sequencing. The predicted ATP6 polypeptide has a 107-amino acid cleavable mitochondrial targeting sequence. The mean hydrophobicity of the protein is decreased in those transmembrane regions that are predicted not to participate directly in proton translocation or in intersubunit contacts with the multimeric ring of c subunits. This is the first example of a mitochondrial protein with more than two transmembrane stretches, directly involved in proton translocation, that is nucleus-encoded.

* This work was supported by the Fogarty International Center at the National Institutes of Health (NIH) Grant TW01176; NHLBI, NIH, Grant HL59646; Consejo Nacional de Ciencia y Tecnologia (CONACyT) Grant 27754N; and Dirección General de Estudios de Personal Académico, Universidad Nacional Autónoma de México (UNAM), Grant IN202598.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequences reported in this paper have been submitted to the DDBJ/GenBankTM/EBI Data Bank with accession numbers AF411119 (atp6 cDNA sequence from C. reinhardtii) and AF411921 (atp6 genomic sequence from C. reinhardtii).

§ Recipient of a fellowship from CONACyT and Dirección General de Posgrado (UNAM) and a graduate student grant from Progama de Apoyo a los Estudios de Posgrado (UNAM).

** Recipient of a fellowship from Dirección General de Posgrado (UNAM).

Dagger Dagger Recipient of a fellowship from CONACyT and Dirección General de Posgrado (UNAM) and a graduate student grant from Progama de Apoyo a los Estudios de Posgrado (UNAM).

§§ To whom correspondence should be addressed: Dept. de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Apartado Postal 70-243, 04510 México D.F., Mexico. Tel.: 52-55-5622-5620; Fax: 52-55-5622-5611; E-mail: dhalphen@ifisiol.unam.mx.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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