Human DNA topoisomerase I-mediated cleavages stimulated by ultraviolet light-induced DNA damage

Annalisa Lanza, Silvia Tornaletti, Carlo Rodolfo, Maria Cristina Scanavini, Antonia M. Pedrini

Research output: Contribution to journalArticlepeer-review

Abstract

DNA topoisomerases have been proposed as the proteins involved in the formation of the DNA-protein cross-links detected after ultraviolet light (UV) irradiation of cellular DNA. This possibility has been investigated by studying the effects of UV-induced DNA damage on human DNA topoisomerase I action. UV lesions impaired the enzyme's ability to relax negatively supercoiled DNA. Decreased relaxation activity correlated with the stimulation of cleavable complexes. Accumulation of clearable complexes resulted from blockage of the rejoining step of the cleavage-religation reaction. Mapping of cleavage sites on the pAT153 genome indicated UV- induced cleavage at discrete positions corresponding to sites stimulated also by the topoisomerase I inhibitor camptothecin, except for one. Subsequent analysis at nucleotide level within the sequence encompassing the UV-specific cleavage site revealed the precise positions of sites stimulated by camptothecin with respect to those specific for UV irradiation. Interestingly, one of the UV-stimulated cleavage sites was formed within a sequence that did not contain dimerized pyrimidines, suggesting transmission of the distortion, caused by photodamage to DNA, into the neighboring sequences. These results support the proposal that DNA structural alterations induced by UV lesions can be sufficient stimulus to induce cross-linking of topoisomerase I to cellular DNA.

Original languageEnglish
Pages (from-to)6978-6986
Number of pages9
JournalJournal of Biological Chemistry
Volume271
Issue number12
DOIs
Publication statusPublished - Mar 22 1996

ASJC Scopus subject areas

  • Biochemistry

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