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

Document Type

Article

Publication Date

1-1-2016

Subject Area

Biosensing Techniques (methods); DNA Polymerase gamma; DNA Replication (genetics); DNA, Mitochondrial (genetics); DNA, Single-Stranded (genetics, metabolism); DNA-Binding Proteins (metabolism); DNA-Directed DNA Polymerase (genetics); Humans; Interferometry (methods); Mitochondria (genetics); Mitochondrial Diseases (genetics); Multienzyme Complexes (genetics)

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.

Publication Title

Methods in molecular biology (Clifton, N.J.)

Volume

1351

First Page

223

Last Page

31

Digital Object Identifier (DOI)

10.1007/978-1-4939-3040-1_17

PubMed ID

26530686

E-ISSN

1940-6029

Language

eng

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