Rolling Circle Replication and Bypass of Damaged Nucleotides

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2023 Feb 22

PMID 36807794

Authors

Josefin M E Forslund

Gorazd Stojkovic

Sjoerd Wanrooij

Affiliations

Soon will be listed here.

Abstract

Faithful mitochondrial DNA (mtDNA) replication is critical for the proper function of the oxidative phosphorylation system. Problems with mtDNA maintenance, such as replication stalling upon encountering DNA damage, impair this vital function and can potentially lead to disease. An in vitro reconstituted mtDNA replication system can be used to investigate how the mtDNA replisome deals with, for example, oxidatively or UV-damaged DNA. In this chapter, we provide a detailed protocol on how to study the bypass of different types of DNA damage using a rolling circle replication assay. The assay takes advantage of purified recombinant proteins and can be adapted to the examination of various aspects of mtDNA maintenance.

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