Analysis of Mitochondrial SSB-DNA Complexes and Their Effects on DNA Polymerase γ Activity by Electron Microscopy and Enzymatic Assays
Document Type
Article
Publication Date
1-1-2021
Subject Area
DNA Polymerase gamma (metabolism); DNA, Single-Stranded (chemistry, metabolism); DNA-Binding Proteins (metabolism); Enzyme Assays; Humans; Microscopy, Electron; Mitochondrial Proteins (metabolism); Molecular Conformation; Nucleoproteins (chemistry)
Abstract
The mitochondrial single-stranded DNA-binding protein (mtSSB) regulates the function of the mitochondrial DNA (mtDNA) replisome. In vitro, mtSSB stimulates the activity of enzymatic components of the replisome, namely mtDNA helicase and DNA polymerase gamma (Pol γ). We have demonstrated that the stimulatory properties of mtSSB result from its ability to organize the single-stranded DNA template in a specific manner. Here we present methods employing electron microscopy and enzymatic assays to characterize and classify the mtSSB-DNA complexes and their effects on the activity of Pol γ.
Publication Title
Methods in molecular biology (Clifton, N.J.)
Volume
2281
First Page
265
Last Page
272
Digital Object Identifier (DOI)
10.1007/978-1-0716-1290-3_16
PubMed ID
33847964
E-ISSN
1940-6029
Language
eng
Citation Information
Bermek, Oya and Ciesielski, Grzegorz L., "Analysis of Mitochondrial SSB-DNA Complexes and Their Effects on DNA Polymerase γ Activity by Electron Microscopy and Enzymatic Assays" (2021). UNF Faculty Research and Scholarship. 3280.
https://digitalcommons.unf.edu/unf_faculty_publications/3280