title: DNA repair synthesis and ligation affect the processing of excised oligonucleotides generated by human nucleotide excision repair.
abstract: Ultraviolet (UV) photoproducts are removed from genomic DNA by dual incisions in humans in the form of 24- to 32-nucleotide-long oligomers (canonical 30-mers) by the nucleotide excision repair system. How the small, excised, damage-containing DNA oligonucleotides (sedDNAs) are processed in cells following the dual incision event is not known. Here, we demonstrate that sedDNAs are localized to the nucleus in two biochemically distinct forms, which include chromatin-associated, transcription factor II H-bound complexes and more readily solubilized, RPA-bound complexes. Because the nuclear mobility and repair functions of transcription factor II H and RPA are influenced by post-incision gap-filling events, we examined how DNA repair synthesis and DNA ligation affect sedDNA processing. We found that although these gap filling activities are not essential for the dual incision/sedDNA generation event per se, the inhibition of DNA repair synthesis and ligation is associated with a decrease in UV photoproduct removal rate and an accumulation of RPA-sedDNA complexes in the cell. These findings indicate that sedDNA processing and association with repair proteins following the dual incisions may be tightly coordinated with gap filling during nucleotide excision repair in vivo.
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599 and.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Kemp
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599 and.
authors.forename: Shobhan
authors.initials: S
authors.lastname: Gaddameedhi
authors.affiliation: Center for Bioanalysis, Department of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon 305-340, South Korea.
authors.forename: Jun-Hyuk
authors.initials: JH
authors.lastname: Choi
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599 and.
authors.forename: Jinchuan
authors.initials: J
authors.lastname: Hu
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599 and. Electronic address: aziz_sancar@med.unc.edu.
authors.forename: Aziz
authors.initials: A
authors.lastname: Sancar
mesh_terms: D059447:Cell Cycle Checkpoints; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D053837:DNA Cleavage; D004260:DNA Repair; D006801:Humans; D011089:Polydeoxyribonucleotides / Q000378:metabolism*; D051716:Replication Protein A / Q000378:metabolism; D051758:Transcription Factor TFIIH / Q000378:metabolism
chemical_list: D011089:Polydeoxyribonucleotides; C497167:RPA1 protein, human; D051716:Replication Protein A; D051758:Transcription Factor TFIIH; D004247:DNA
keywords: DNA Damage; DNA Damage Response; DNA Repair; DNA-Protein Interaction; Nucleotide Excision Repair; Subcellular Fractionation
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