Mass Spectrometry-based Assays for Examining the Repair of Bulky DNA Lesions Induced by Reactive Oxygen Species

摘要

Endogenous metabolism and exposure to environmental agents can lead to cellular production of reactive oxygen species (ROS), which can cause damage to DNA. Although substantial repair studies have been conducted for the ROS-induced singlenucleobase lesions, not much is known about how oxidatively induced bulky DNA lesions are repaired in mammalian cells. Here, we describe two MS-based methods for assessing the repair of the latter type of lesions in mammalian cells. The first method relies on the use of LC-MS/MS/MS for the quantification of these lesions in tissues of animals that are repair-proficient or deficient in certain repair proteins. In the second method, we combine shuttle vector technology with MS to ask how DNA lesions perturb DNA transcription in mammalian cells that are proficient or deficient in DNA repair.