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Purification, Separation, and Identification of the Human mtDNA Polymerase With and Without Its Accessory Subunit

  • Protocol
Mitochondrial DNA

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 197))

Abstract

The human mitochondrial genome encodes a variety of genes required for oxidative phosphorylation, and loss of these essential gene functions induces a multitude of severe metabolic disorders (1–4). Mitochondrial DNA (mtDNA) is replicated by the nuclear encoded DNA polymerase γ (5), and pol γ is the only replicative eukaryotic DNA polymerase to display sensitivity to a wide range of antiviral nucleotide analogs (6–12). Consequently, long-term treatment of patients with antiviral nucleotide analogs such as zidovudine (AZT), zalcitabine (ddC), and didanosine (ddI) induces a characteristic collection of mitochondrial dysfunctions that mimics mitochondrial genetic diseases (13–16). To aid investigations into the properties and subunit composition of this important enzyme, we present a simple biochemical assay to distinguish two forms of pol γ and a purification scheme to resolve them.

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Authors and Affiliations

  1. Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC

    Matthew J. Longley & William C. Copeland

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  1. Matthew J. Longley
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  2. William C. Copeland
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Editors and Affiliations

  1. Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC

    William C. Copeland

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© 2002 Humana Press Inc.

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Longley, M.J., Copeland, W.C. (2002). Purification, Separation, and Identification of the Human mtDNA Polymerase With and Without Its Accessory Subunit. In: Copeland, W.C. (eds) Mitochondrial DNA. Methods in Molecular Biology™, vol 197. Humana Press. https://doi.org/10.1385/1-59259-284-8:245

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  • DOI: https://doi.org/10.1385/1-59259-284-8:245

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-972-8

  • Online ISBN: 978-1-59259-284-5

  • eBook Packages: Springer Protocols

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