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Papers In Press, published online ahead of print December 14, 2001
Departamento de Genetica Molecular, Instituto de Fisiología Celular, UNAM, Institute of Cellular Physiology, UNAM, Mexico City, D.F. 04510
Corresponding Author: dhalphen{at}ifisiol.unam.mx
The atp6 gene, encoding the ATP6 subunit of the mitochondrial 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. We investigated the possibility that ATP6 is nuclear-encoded in this green alga. Analysis of C. reinhardtii ESTs revealed three overlapping sequences that encoded a protein with similarity to ATP6 proteins. PCR and 5'- and 3'- RACE, using primers designed from the EST sequences, were used to obtain the complete cDNA and genomic sequences of C. reinhardtii atp6. The atp6 gene was shown to be located in the nucleus and to have characteristics of a nuclear-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 mitochondria by N-terminal sequencing of the subunits of this protein complex. The predicted ATP6 polypeptide has a 107 amino acid cleavable mitochondrial targeting sequence and exhibits diminished physical constraints for import into mitochondria. The mean hydrophobicity of the protein is decreased in those transmembrane regions that are predicted not to participate directly in proton translocation or in inter-subunit 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 nuclear encoded.
J. Biol. Chem, 10.1074/jbc.M109993200
Submitted on October 16, 2001
Revised on December 14, 2001
Accepted on December 14, 2001
The typically mtDNA-encoded ATP6 subunit of the mitochondrial F1F0-ATPase is encoded by a nuclear gene in Chlamydomonas reinhardtii
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