Contribution of DNA unwrapping from histone octamers to the repair of oxidatively damaged DNA in nucleosomes.
Reactive oxygen species generate ~20,000 oxidative lesions in the DNA of every cell, every day. Most of these lesions are located within nucleosomes, which package DNA in chromatin and impede base excision repair (BER). We demonstrated previously that periodic, spontaneous partial unwrapping of DNA from the underlying histone octamer enables BER enzymes to bind to oxidative lesions that would otherwise be sterically inaccessible. In the present study, we asked if these periodic DNA unwrapping events are frequent enough to account for the estimated rates of BER in vivo. We measured rates of excision of oxidative lesions from sites in nucleosomes that are accessible only during unwrapping episodes. Using reaction conditions appropriate for presteady-state kinetic analyses, we derived lesion exposure rates for both 601 and 5S rDNA-based nucleosomes. Although DNA unwrapping-mediated exposure of a lesion ~16NT from the nucleosome edge occurred ~7-8 times per minute, exposure rates fell dramatically for lesions located 10 or more NT further in from the nucleosome edge. The rates likely are too low to account for observed rates of BER in cells. Thus, chromatin remodeling, either BER-specific or that associated with transcription, replication, or other DNA repair processes, probably contributes to efficient BER in vivo.
Department of Microbiology and Molecular Genetics, University of Vermont, Burlington, VT 05405, USA.
Robyn L
RL
Maher
Amalthiya
A
Prasad
Olga
O
Rizvanova
Susan S
SS
Wallace
David S
DS
Pederson
D042002:Chromatin Assembly and Disassembly; D004249:DNA Damage; D045647:DNA Glycosylases / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair / Q000235:genetics* / Q000502:physiology; D004275:DNA, Ribosomal / Q000235:genetics; D006657:Histones / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009707:Nucleosomes / Q000235:genetics* / Q000378:metabolism; D018384:Oxidative Stress; D013941:Thymine / Q000031:analogs & derivatives / Q000378:metabolism
D004275:DNA, Ribosomal; D006657:Histones; D009707:Nucleosomes; C029389:thymine glycol; D045647:DNA Glycosylases; D013941:Thymine
Base excision repair; Chromatin; DNA unwrapping kinetics; Human NTH1 kinetics; Nucleosome dynamics