A significant UV-induced photoproduct of DNA, cyclobutane pyrimidine dimer (CPD), occurs by dimerization of adjacent pyrimidine bases. This photoproduct forms lesions on DNA, which can be repaired by photolyases. This research will utilize resonance Raman spectroscopy to study the changes induced by binding Escherichia coli photolyase to damaged DNA. A poly dT 10-mer will be irradiated with UV light to form CPDs between adjacent thymidines (TT). The damaged poly dT 10-mer will be combined with photolyase for resonance Raman analysis. An oligonucleotide (5'-CGGCATTACGGC-3') will be irradiated, using acetophenone as a photosensitizer, to form TT between the central thymidines. These damaged oligonucleotide strands can then be purified using HPLC and combined with photolyase for analysis. The Raman spectra of photolyase bound to both ssDNA and dsDNA will be compared to the spectrum of free photolyase to determine the impact of binding on the interaction near the active site of the enzyme.

Spectroscopic study of the interaction of <i>Escherichia coli</i> photolyase with UV-damaged DNA

244th National Meeting (2012)

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Spectroscopic study of the interaction of Escherichia coli photolyase with UV-damaged DNA