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000276790 1001_ $$aSchmalz, Michael$$b0
000276790 245__ $$aDissection of DNA damage and repair pathways in live cells by femtosecond laser microirradiation and free-electron modeling.
000276790 260__ $$aWashington, DC$$bNational Acad. of Sciences$$c2023
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000276790 520__ $$aUnderstanding and predicting the outcome of the interaction of light with DNA has a significant impact on the study of DNA repair and radiotherapy. We report on a combination of femtosecond pulsed laser microirradiation at different wavelengths, quantitative imaging, and numerical modeling that yields a comprehensive picture of photon-mediated and free-electron-mediated DNA damage pathways in live cells. Laser irradiation was performed under highly standardized conditions at four wavelengths between 515 nm and 1,030 nm, enabling to study two-photon photochemical and free-electron-mediated DNA damage in situ. We quantitatively assessed cyclobutane pyrimidine dimer (CPD) and γH2AX-specific immunofluorescence signals to calibrate the damage threshold dose at these wavelengths and performed a comparative analysis of the recruitment of DNA repair factors xeroderma pigmentosum complementation group C (XPC) and Nijmegen breakage syndrome 1 (Nbs1). Our results show that two-photon-induced photochemical CPD generation dominates at 515 nm, while electron-mediated damage dominates at wavelengths ≥620 nm. The recruitment analysis revealed a cross talk between nucleotide excision and homologous recombination DNA repair pathways at 515 nm. Numerical simulations predicted electron densities and electron energy spectra, which govern the yield functions of a variety of direct electron-mediated DNA damage pathways and of indirect damage by •OH radicals resulting from laser and electron interactions with water. Combining these data with information on free electron-DNA interactions gained in artificial systems, we provide a conceptual framework for the interpretation of the wavelength dependence of laser-induced DNA damage that may guide the selection of irradiation parameters in studies and applications that require the selective induction of DNA lesions.
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000276790 650_7 $$2Other$$aDNA strand breaks
000276790 650_7 $$2Other$$anonlinear photodamage
000276790 650_7 $$2Other$$aoxidative DNA damage
000276790 650_7 $$2Other$$areductive DNA damage
000276790 650_7 $$2Other$$awavelength selectivity
000276790 7001_ $$00000-0002-8325-1627$$aLiang, Xiao-Xuan$$b1
000276790 7001_ $$aWieser, Ines$$b2
000276790 7001_ $$00009-0000-7452-5326$$aGruschel, Caroline$$b3
000276790 7001_ $$aMuskalla, Lukas$$b4
000276790 7001_ $$aStöckl, Martin Thomas$$b5
000276790 7001_ $$00000-0002-9397-8475$$aNitschke, Roland$$b6
000276790 7001_ $$00000-0002-7843-5966$$aLinz, Norbert$$b7
000276790 7001_ $$00000-0002-9847-257X$$aLeitenstorfer, Alfred$$b8
000276790 7001_ $$00000-0002-4371-9037$$aVogel, Alfred$$b9
000276790 7001_ $$00000-0002-5567-8690$$aFerrando-May, Elisa$$b10$$eLast author
000276790 773__ $$0PERI:(DE-600)1461794-8$$a10.1073/pnas.2220132120$$gVol. 120, no. 25, p. e2220132120$$n25$$pe2220132120$$tProceedings of the National Academy of Sciences of the United States of America$$v120$$x0027-8424$$y2023
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