Mapping atypical UV photoproducts in vitro and across the S. cerevisiae genome

Kaitlynne A Bohm; Smitha Sivapragasam; John J Wyrick

doi:10.1016/j.xpro.2021.101059

Mapping atypical UV photoproducts in vitro and across the S. cerevisiae genome

STAR Protoc. 2021 Dec 22;3(1):101059. doi: 10.1016/j.xpro.2021.101059. eCollection 2022 Mar 18.

Authors

Kaitlynne A Bohm¹, Smitha Sivapragasam¹, John J Wyrick^{1

2}

Affiliations

¹ School of Molecular Biosciences, Washington State University, Pullman, WA 99164, USA.
² Center for Reproductive Biology, Washington State University, Pullman, WA 99164, USA.

Abstract

Exposure to ultraviolet (UV) light induces DNA damage, predominantly cyclobutane pyrimidine dimers (CPD) and 6,4-photoproducts (6,4-PP), as well as rare, atypical photoproducts at thymidine-adenine (TA) sequences. We have recently shown 'TA' photoproducts are induced in UV-irradiated oligonucleotides and across the budding yeast genome. Here, we describe a protocol for mapping atypical 'TA' photoproducts in vitro and in vivo. This protocol overcomes the technical challenges involved in accurately mapping such rare photoproducts by using ultraviolet damage endonuclease (UVDE) enzymes. For complete details on the use and execution of this protocol, please refer to Laughery et al. (2020).

Keywords: Bioinformatics; Model Organisms; Molecular Biology; Sequence analysis; Sequencing.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

DNA Damage / genetics
DNA Repair* / genetics
Pyrimidine Dimers
Saccharomyces cerevisiae* / genetics
Ultraviolet Rays / adverse effects

Substances

Pyrimidine Dimers

Abstract

Publication types

MeSH terms

Substances

Grants and funding