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J. Biol. Chem., Vol. 280, Issue 5, 3242-3250, February 4, 2005

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Bidirectional Replication Initiates at Sites Throughout the Mitochondrial Genome of Birds^*

Aurelio Reyes¶, Ming Yao Yang, Mark Bowmaker, and Ian J. Holt||

From the Medical Research Council Dunn Human Nutrition Unit, Wellcome Trust/MRC Building, Hills Road Cambridge, CB2 2XY, United Kingdom and the Sezione di Bioinformatica e Genomica di Bari, Istituto Tecnologie Biomediche, CNR, Via Amendola 168/5, 70126 Bari, Italy

Analysis of mitochondrial replication intermediates of Gallus gallus on fork-direction gels indicates that replication occurs in both directions around circular mitochondrial DNA. This finding was corroborated by a study of chick mitochondrial DNA on standard neutral two-dimensional agarose gels, which yielded archetypal initiation arcs in fragments covering the entire genome. There was, however, considerable variation in initiation arc intensity. The majority of initiation events map to regions flanking the major non-coding region, in particular the NADH dehydrogenase subunit 6 (ND6) gene. Initiation point mapping of the ND6 gene identified prominent free 5' ends of DNA, which are candidate start sites for DNA synthesis. Therefore we propose that the initiation zone of G. gallus mitochondrial DNA encompasses most, if not all, of the genome, with preferred initiation sites in regions flanking the major non-coding region. Comparison with mammals suggests a common mechanism of initiation of mitochondrial DNA replication in higher vertebrates.

Received for publication, October 20, 2004 , and in revised form, November 19, 2004.

^* 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 information.

^¶ Recipient of an EMBO short-term fellowship (April–July 2001, ASTF9784) and a Royal Society Travel Fellowship (September 2002–August 2003) and is currently supported by the European Union sixth Framework Programme for Research, Priority 1 "Life sciences, genomics and biotechnology for health" contract number LSHM-CT-2004-503116, and the European Commission (QLG1-CT-2001-00966).

^|| To whom correspondence should be addressed. Tel.: 44-12-23-25-28-40; Fax: 44-12-23-25-28-45; E-mail: holt{at}mrc-dunn.cam.ac.uk.

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