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@ARTICLE{Schmalz:276790,
author = {M. Schmalz and X.-X. Liang and I. Wieser and C. Gruschel
and L. Muskalla and M. T. Stöckl and R. Nitschke and N.
Linz and A. Leitenstorfer and A. Vogel and E.
Ferrando-May$^*$},
title = {{D}issection of {DNA} damage and repair pathways in live
cells by femtosecond laser microirradiation and
free-electron modeling.},
journal = {Proceedings of the National Academy of Sciences of the
United States of America},
volume = {120},
number = {25},
issn = {0027-8424},
address = {Washington, DC},
publisher = {National Acad. of Sciences},
reportid = {DKFZ-2023-01165},
pages = {e2220132120},
year = {2023},
note = {#LA:W650#},
abstract = {Understanding 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.},
keywords = {DNA strand breaks (Other) / nonlinear photodamage (Other) /
oxidative DNA damage (Other) / reductive DNA damage (Other)
/ wavelength selectivity (Other)},
cin = {W650},
ddc = {500},
cid = {I:(DE-He78)W650-20160331},
pnm = {312 - Funktionelle und strukturelle Genomforschung
(POF4-312)},
pid = {G:(DE-HGF)POF4-312},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:37307476},
doi = {10.1073/pnas.2220132120},
url = {https://inrepo02.dkfz.de/record/276790},
}