Chapter title |
Mitochondrial DNA
|
---|---|
Chapter number | 2 |
Book title |
Mitochondrial DNA
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3040-1_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3039-5, 978-1-4939-3040-1
|
Authors |
Copeland, William C, Kasiviswanathan, Rajesh, Longley, Matthew J, William C. Copeland, Rajesh Kasiviswanathan, Matthew J. Longley |
Abstract |
Mitochondrial DNA is replicated by the nuclear-encoded DNA polymerase γ (pol γ) which is composed of a single 140 kDa catalytic subunit and a dimeric 55 kDa accessory subunit. Mitochondrial DNA is vulnerable to various forms of damage, including several types of oxidative lesions, UV-induced photoproducts, chemical adducts from environmental sources, as well as alkylation and inter-strand cross-links from chemotherapy agents. Although many of these lesions block DNA replication, pol γ can bypass some lesions by nucleotide incorporation opposite a template lesion and further extension of the DNA primer past the lesion. This process of translesion synthesis (TLS) by pol γ can occur in either an error-free or an error-prone manner. Assessment of TLS requires extensive analysis of oligonucleotide substrates and replication products by denaturing polyacrylamide sequencing gels. This chapter presents protocols for the analysis of translesion DNA synthesis. |
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