Nucleic Acids Research, 2003, Vol. 31, No. 11 2786-2794
© 2003 Oxford University Press
UVA-induced cyclobutane pyrimidine dimers form predominantly at thymine–thymine dipyrimidines and correlate with the mutation spectrum in rodent cells
1 Division of Pathology, Department of Medical Biology, Faculty of Medicine, Laval University and Unité de Recherche en Génétique Humaine et Moléculaire, Research Center, Hôpital Saint-François d’Assise, Centre Hospitalier Universitaire de Québec, Québec, QC G1L 3L5, Canada, 2 Faculty of Medicine, University of Montreal and Centre de Recherche Guy-Bernier, Hôpital Maisonneuve-Rosemont, Montréal, QC H1T 2M4, Canada and 3 CNRS UMR 2027, Institut Curie, Bât. 110, Centre Universitaire, F-91405 Orsay, France
*To whom correspondence should be addressed. Tel: +33 1 69 86 71 87; Fax: +33 1 69 86 94 29; Email: evelyne.sage{at}curie.u-psud.fr
Ligation-mediated PCR was employed to quantify cyclobutane pyrimidine dimer (CPD) formation at nucleotide resolution along exon 2 of the adenine phosphoribosyltransferase (aprt) locus in Chinese hamster ovary (CHO) cells following irradiation with either UVA (340–400 nm), UVB (295–320 nm), UVC (254 nm) or simulated sunlight (SSL; 
> 295 nm). The resulting DNA damage spectrum for each wavelength region was then aligned with the corresponding mutational spectrum generated previously in the same genetic target. The DNA sequence specificities of CPD formation induced by UVC, UVB or SSL were very similar, i.e., in each case the overall relative proportion of this photoproduct forming at TT, TC, CT and CC sites was 
28, 26, 16 and 30%, respectively. Furthermore, a clear correspondence was noted between the precise locations of CPD damage hotspots, and of ‘UV signature’ mutational hotspots consisting primarily of C
T and CCTT transitions within pyrimidine runs. However, following UVA exposure, in strong contrast to the above situation for UVC, UVB or SSL, CPDs were generated much more frequently at TT sites than at TC, CT or CC sites (57% versus 18, 11 and 14%, respectively). This CPD deposition pattern correlates well with the strikingly high proportion of mutations recovered opposite TT dipyrimidines in UVA- irradiated CHO cells. Our results directly implicate the CPD as a major promutagenic DNA photoproduct induced specifically by UVA in rodent cells.
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