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Mitochondria pp 193–206Cite as

In Vitro Analysis of the Yeast Mitochondrial RNA Polymerase

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 372))

Abstract

Understanding the details of how genetic information is expressed from the separate mitochondrial genome requires a detailed description of the properties of the mitochondrial RNA polymerase. This nuclear-encoded enzyme is necessary and sufficient for the transcription of all mitochondrially encoded genes. Mitochondria from yeast to humans use a single-polypeptide catalytic RNA polymerase related to enzymes from bacteriophage. They also require separable transcription factors necessary for initiation at promoter sequences on the mitochondrial DNA template. It has recently become possible to work with highly purified, recombinant forms of the mitochondrial RNA polymerase subunits from yeast. This chapter describes detailed protocols for working in vitro with this purified enzyme in transcription reactions. These assays are critical for elucidating the nature of a mitochondrial promoter and for understanding how the mitochondrial RNA polymerase recognizes these DNA sequences and selectively initiates the transcription cycle, resulting in discrete transcripts.

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Author information

Authors and Affiliations

  1. Department of Biochemistry, University of Utah, Salt Lake City, UT

    Elizabeth A. Amiott

  2. Department of Biochemistry and Molecular Genetics, University of Colorado at Denver and Health Sciences Center, Aurora, CO

    Judith A. Jaehning

Authors
  1. Elizabeth A. Amiott
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  2. Judith A. Jaehning
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Editor information

Editors and Affiliations

  1. Department für Biologie I, Botanik, Ludwig-Maximilians-Universität München, München, Germany

    Dario Leister

  2. Zellbiologie, Universität Kaiserslautern, Kaiserslautern, Germany

    Johannes M. Herrmann

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© 2007 Humana Press Inc., Totowa, NJ

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Amiott, E.A., Jaehning, J.A. (2007). In Vitro Analysis of the Yeast Mitochondrial RNA Polymerase. In: Leister, D., Herrmann, J.M. (eds) Mitochondria. Methods in Molecular Biology™, vol 372. Humana Press. https://doi.org/10.1007/978-1-59745-365-3_14

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  • DOI: https://doi.org/10.1007/978-1-59745-365-3_14

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-667-2

  • Online ISBN: 978-1-59745-365-3

  • eBook Packages: Springer Protocols

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