Analysis of Base Excision and Single-Strand Break Repair Activities in Trypanosomatid Extracts

Daria Kania, Michael Ginger, Sarah Allinson

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Cellular DNA is inherently unstable, subject to both spontaneous hydrolysis and attack by a range of exogenous and endogenous chemicals as well as physical agents such as ionizing and ultraviolet radiation. For parasitic protists, where an inoculum of infectious parasites is typically small and natural infections are often chronic with low parasitemia, they are also vulnerable to DNA damaging agents arising from innate immune defenses. The majority of DNA damage consists of relatively minor changes to the primary structure of the DNA, such as base deamination, oxidation, or alkylation and scission of the phosphodiester backbone. Yet these small changes can have serious consequences, often being mutagenic or cytotoxic. Cells have therefore evolved efficient mechanisms to repair such damage, with base excision and single strand break repair playing the primary role here. In this chapter we describe a method for analyzing the activity from cell extracts of various enzymes involved in the base excision and single strand break repair pathways of trypanosomatid parasites.
Original languageEnglish
Title of host publicationTrypanosomatids
Subtitle of host publicationMethods and Protocols
EditorsPaul A. M. Michels, Michael L. Ginger, Dan Zilberstein
PublisherHumana Press
Chapter22
Pages353-364
Number of pages12
Volume2116
ISBN (Electronic)9781071602942
ISBN (Print)9781071602935
DOIs
Publication statusPublished - 28 Mar 2020

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
Volume2116
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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  • Cite this

    Kania, D., Ginger, M., & Allinson, S. (2020). Analysis of Base Excision and Single-Strand Break Repair Activities in Trypanosomatid Extracts. In P. A. M. Michels, M. L. Ginger, & D. Zilberstein (Eds.), Trypanosomatids: Methods and Protocols (Vol. 2116, pp. 353-364). (Methods in Molecular Biology; Vol. 2116). Humana Press. https://doi.org/10.1007/978-1-0716-0294-2_22