Study of UV-induced DNA Repair Factor Recruitment: Kinetics and Dynamics

Sarah Sertic; Stefania Roma; Paolo Plevani; Federico Lazzaro; Marco Muzi-Falconi

doi:10.1007/978-1-4939-7306-4_8

Study of UV-induced DNA Repair Factor Recruitment: Kinetics and Dynamics

Methods Mol Biol. 2018:1672:101-105. doi: 10.1007/978-1-4939-7306-4_8.

Authors

Sarah Sertic¹, Stefania Roma¹, Paolo Plevani¹, Federico Lazzaro¹, Marco Muzi-Falconi²

Affiliations

¹ Dipartimento di Bioscienze, Università degli Studi di Milano, Via Celoria 26, Milano, 20133, Italy.
² Dipartimento di Bioscienze, Università degli Studi di Milano, Via Celoria 26, Milano, 20133, Italy. marco.muzifalconi@unimi.it.

PMID: 29043619
DOI: 10.1007/978-1-4939-7306-4_8

Abstract

The local UV irradiation technique enables detection, kinetic measurements of recruitment, and quantification of DNA Damage Response (DDR) proteins at the site of UV-induced DNA damage.Using Isopore filters with high density pores of a broad range of sizes, it is possible to UV irradiate and damage only a very small portion of the nucleus of a cell by letting UV light pass only through the pores. Immunofluorescent analyses of modified DNA nucleotides, proteins, or fluorescently tagged versions of target factors can be used as markers to label and study UV-induced lesions and their repair.

Keywords: Immunofluorescence; NER; Protein recruitment; Sequential assembly; UV-induced DNA damage.

MeSH terms

DNA Damage / radiation effects*
DNA Repair / radiation effects*
DNA-Binding Proteins / metabolism*
Fluorescent Antibody Technique
Microscopy, Fluorescence
Protein Binding
Ultraviolet Rays*

Substances

DNA-Binding Proteins