DNA repair synthesis and ligation affect the processing of excised oligonucleotides generated by human nucleotide excision repair

Michael G Kemp; Shobhan Gaddameedhi; Jun-Hyuk Choi; Jinchuan Hu; Aziz Sancar

doi:10.1074/jbc.M114.597088

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DNA repair synthesis and ligation affect the processing of excised oligonucleotides generated by human nucleotide excision repair

Journal article

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DNA repair synthesis and ligation affect the processing of excised oligonucleotides generated by human nucleotide excision repair

Michael G Kemp, Shobhan Gaddameedhi, Jun-Hyuk Choi, Jinchuan Hu and Aziz Sancar

The Journal of biological chemistry, Vol.289(38), pp.26574-26583

09/19/2014

DOI: https://doi.org/10.1074/jbc.M114.597088

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https://hdl.handle.net/2376/107966

PMCID: PMC4176238

PMID: 25107903

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Abstract

Polydeoxyribonucleotides - metabolism

Cell Cycle Checkpoints

Cell Nucleus - metabolism

DNA Repair

Humans

Transcription Factor TFIIH - metabolism

Cell Line, Tumor

DNA - biosynthesis

Replication Protein A - metabolism

DNA Cleavage

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.

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Title: DNA repair synthesis and ligation affect the processing of excised oligonucleotides generated by human nucleotide excision repair
Creators: Michael G Kemp - Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599 and
Shobhan Gaddameedhi - Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599 and
Jun-Hyuk Choi - Center for Bioanalysis, Department of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon 305-340, South Korea
Jinchuan Hu - Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599 and
Aziz Sancar - 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
Publication Details: The Journal of biological chemistry, Vol.289(38), pp.26574-26583
Academic Unit: UNKNOWN
Publisher: United States
Grant note: GM32833 / NIGMS NIH HHS P30 CA016086 / NCI NIH HHS R00 ES022640 / NIEHS NIH HHS K99 ES022640 / NIEHS NIH HHS K99ES022640 / NIEHS NIH HHS R01 GM032833 / NIGMS NIH HHS
Identifiers: 99900546836401842
Language: English
Resource Type: Journal article