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© 1994 Oxford University Press

research-article

Immunochemical detection of sequence-specific modifications to DNA induced by UV light

Karl E. Herbert, Nalini Mistry, Helen R. Griffiths and Joseph Lunec

Division of Chemical Pathology Centre for Mechanisms of Human Toxicity, P0 Box 138, University of Leicester, Lancaster Road, Leicester LEI 9HN and Clinical Sciences. Glenfield Hospital NHS Trust, Groby Road, Leicester LE3 9QP, UK

Sequence specificity of antibodies to UV-damaged DNA has not been described previously. The antisera Investigated here were specific for UV-modifled DNA and were absolutely dependent upon the presence of thymine residues. Using a series of oligonucleotides in competition ELISA, Increased Inhibition was observed with Increasing chain length of UV-polythymldylate. A minimum of three adja cent thymlnes was required for effective inhibition; alone, dimers of thymine were poor antigens. Although UV Irradiated poly(dC) was not anfigenic, cytosines could partially replace thymines within the smallest effective epitope (T-T-T) with a high degree of sequence specificity, not previously described. The main epitope induced by UV was formed from adjacent thymines and either a 3' or a 5' pyrhnidlne.


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