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Pyrimidine (6–4) Pyrimidone Photoproduct Mapping After Sublethal UVC Doses: Nucleotide Resolution Using Terminal Transferase‐dependent PCR
Author(s) -
Rochette Patrick J.,
Bastien Nathalie,
Todo Takeshi,
Drouin Régen
Publication year - 2006
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1562/2004-12-01-ra-390
Subject(s) - pyrimidine dimer , nucleotide , photolyase , genomic dna , microbiology and biotechnology , dna , cyclobutane , biology , chemistry , dna damage , dna repair , genetics , gene , ring (chemistry) , organic chemistry
UVC irradiation of genomic DNA induces two main types of potentially mutagenic base modifications: cyclobutane pyrimidine dimers (CPDs) and the less frequent (15–30% of CPD levels) pyrimidine (6–4) pyrimidone photoproducts (6–4PP). Ligation‐mediated PCR (LMPCR), a genomic sequencing technique, allows CPD mapping at nucleotide resolution following irradiation with sublethal doses of UVB or UVC for most cell types. In contrast, a dose of 80 J/m 2 of UVC that is lethal for the majority of cell types is necessary to map 6–4PP by the LMPCR technique. This compromises the use of LMPCR to study the repair of 6–4PP. To date, no other techniques have been developed to study 6–4PP repair at nucleotide resolution. We have therefore adapted a recently developed technique for the mapping of 6–4PP: terminal transferase‐dependent PCR (TDPCR). TDPCR is in many ways similar to LMPCR. This technique is more sensitive and allows the mapping of 6–4PP at UVC doses as low as 10 J/m 2 in genomic DNA and in living cells.