{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Lan L"],"funding":["NIA NIH HHS","NIEHS NIH HHS","NCI NIH HHS","NIGMS NIH HHS"],"pagination":["2330-45"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC3936713"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["42(4)"],"pubmed_abstract":["Reactive oxygen species (ROS)-induced DNA damage is repaired by the base excision repair pathway. However, the effect of chromatin structure on BER protein recruitment to DNA damage sites in living cells is poorly understood. To address this problem, we developed a method to specifically produce ROS-induced DNA damage by fusing KillerRed (KR), a light-stimulated ROS-inducer, to a tet-repressor (tetR-KR) or a transcription activator (TA-KR). TetR-KR or TA-KR, bound to a TRE cassette (∼ 90 kb) integrated at a defined genomic locus in U2OS cells, was used to induce ROS damage in hetero- or euchromatin, respectively. We found that DNA glycosylases were efficiently recruited to DNA damage in heterochromatin, as well as in euchromatin. PARP1 was recruited to DNA damage within condensed chromatin more efficiently than in active chromatin. In contrast, recruitment of FEN1 was highly enriched at sites of DNA damage within active chromatin in a PCNA- and transcription activation-dependent manner. These results indicate that oxidative DNA damage is differentially processed within hetero or euchromatin."],"journal":["Nucleic acids research"],"pubmed_title":["Novel method for site-specific induction of oxidative DNA damage reveals differences in recruitment of repair proteins to heterochromatin and euchromatin."],"pmcid":["PMC3936713"],"funding_grant_id":["AG045545-01","P30 CA047904","GM099213","GM103529","ES019498","ES019566","GM087798","R21 AG045545","R01 ES019566","CA148629","R01 CA148629"],"pubmed_authors":["Sun L","Bruchez M","Hsieh CL","Lan L","Nakajima S","Yasui A","Van Houten B","Wei L","Sobol RW","Levine AS"],"additional_accession":[]},"is_claimable":false,"name":"Novel method for site-specific induction of oxidative DNA damage reveals differences in recruitment of repair proteins to heterochromatin and euchromatin.","description":"Reactive oxygen species (ROS)-induced DNA damage is repaired by the base excision repair pathway. However, the effect of chromatin structure on BER protein recruitment to DNA damage sites in living cells is poorly understood. To address this problem, we developed a method to specifically produce ROS-induced DNA damage by fusing KillerRed (KR), a light-stimulated ROS-inducer, to a tet-repressor (tetR-KR) or a transcription activator (TA-KR). TetR-KR or TA-KR, bound to a TRE cassette (∼ 90 kb) integrated at a defined genomic locus in U2OS cells, was used to induce ROS damage in hetero- or euchromatin, respectively. We found that DNA glycosylases were efficiently recruited to DNA damage in heterochromatin, as well as in euchromatin. PARP1 was recruited to DNA damage within condensed chromatin more efficiently than in active chromatin. In contrast, recruitment of FEN1 was highly enriched at sites of DNA damage within active chromatin in a PCNA- and transcription activation-dependent manner. These results indicate that oxidative DNA damage is differentially processed within hetero or euchromatin.","dates":{"release":"2014-01-01T00:00:00Z","publication":"2014 Feb","modification":"2025-05-18T12:47:42.623Z","creation":"2025-05-18T12:47:42.623Z"},"accession":"S-EPMC3936713","cross_references":{"pubmed":["24293652"],"doi":["10.1093/nar/gkt1233"]}}