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Biolayer Interferometry: A Novel Method to Elucidate Protein–Protein and Protein–DNA Interactions in the Mitochondrial DNA Replisome

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Mitochondrial DNA

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1351))

Abstract

A lack of effective treatment for mitochondrial diseases prompts scientists to investigate the molecular processes that underlie their development. The major cause of mitochondrial diseases is dysfunction of the sole mitochondrial DNA polymerase, DNA polymerase γ (Pol γ). The development of treatment strategies will require a detailed characterization of the molecular properties of Pol γ. A novel technique, biolayer interferometry, allows one to monitor molecular interactions in real time, thus providing an insight into the kinetics of the process. Here, we present an application of the biolayer interferometry technique to characterize the fundamental reactions that Pol γ undergoes during the initiation phase of mitochondrial DNA replication: holoenzyme formation and binding to the primer-template.

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Acknowledgments

This work was supported by grant GM45295 from the National Institutes of Health, and funds from the University of Tampere to L.S.K. G.C. was supported in part by Biocenter Finland. V.H. was supported by the Academy of Finland (grants 136288 and 273192). We acknowledge infrastructure support from Biocenter Finland.

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Correspondence to Laurie S. Kaguni .

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Ciesielski, G.L., Hytönen, V.P., Kaguni, L.S. (2016). Biolayer Interferometry: A Novel Method to Elucidate Protein–Protein and Protein–DNA Interactions in the Mitochondrial DNA Replisome. In: McKenzie, M. (eds) Mitochondrial DNA. Methods in Molecular Biology, vol 1351. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3040-1_17

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  • DOI: https://doi.org/10.1007/978-1-4939-3040-1_17

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3039-5

  • Online ISBN: 978-1-4939-3040-1

  • eBook Packages: Springer Protocols

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