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/m2 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/m2 in genomic DNA and in living cells.
How to translate text using browser tools
1 September 2006
Pyrimidine (6-4) Pyrimidone Photoproduct Mapping After Sublethal UVC Doses: Nucleotide Resolution Using Terminal Transferase-dependent PCR
Patrick J. Rochette,
Nathalie Bastien,
Takeshi Todo,
Régen Drouin
ACCESS THE FULL ARTICLE
It is not available for individual sale.
This article is only available to subscribers.
It is not available for individual sale.
It is not available for individual sale.
Photochemistry and Photobiology
Vol. 82 • No. 5
September 2006
Vol. 82 • No. 5
September 2006