Nucleotide excision repair in human cells: fate of the excised oligonucleotide carrying DNA damage in vivo

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

doi:10.1074/jbc.M113.482257

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Nucleotide excision repair in human cells: fate of the excised oligonucleotide carrying DNA damage in vivo

Journal article

Open access

Peer reviewed

Nucleotide excision repair in human cells: fate of the excised oligonucleotide carrying DNA damage in vivo

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

The Journal of biological chemistry, Vol.288(29), pp.20918-20926

07/19/2013

DOI: https://doi.org/10.1074/jbc.M113.482257

Handle:

https://hdl.handle.net/2376/104299

PMCID: PMC3774362

PMID: 23749995

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https://doi.org/10.1074/jbc.M113.482257View

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Abstract

Pyrimidine Dimers - metabolism

Cell Line

Humans

Transcription, Genetic - radiation effects

DNA Repair - radiation effects

Mutation - genetics

DNA-Binding Proteins - metabolism

Oligonucleotides - metabolism

Animals

Ultraviolet Rays

Models, Biological

Transcription Factor TFIIH - metabolism

DNA Damage

Nucleotide excision repair is the sole mechanism for removing the major UV photoproducts from genomic DNA in human cells. In vitro with human cell-free extract or purified excision repair factors, the damage is removed from naked DNA or nucleosomes in the form of 24- to 32-nucleotide-long oligomers (nominal 30-mer) by dual incisions. Whether the DNA damage is removed from chromatin in vivo in a similar manner and what the fate of the excised oligomer was has not been known previously. Here, we demonstrate that dual incisions occur in vivo identical to the in vitro reaction. Further, we show that transcription-coupled repair, which operates in the absence of the XPC protein, also generates the nominal 30-mer in UV-irradiated XP-C mutant cells. Finally, we report that the excised 30-mer is released from the chromatin in complex with the repair factors TFIIH and XPG. Taken together, our results show the congruence of in vivo and in vitro data on nucleotide excision repair in humans.

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Title: Nucleotide excision repair in human cells: fate of the excised oligonucleotide carrying DNA damage in vivo
Creators: Jinchuan Hu - From the Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599-7260 and
Jun-Hyuk Choi - From the Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599-7260 and; the Center for Bioanalysis, Department of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon 305-340, South Korea
Shobhan Gaddameedhi - From the Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599-7260 and
Michael G Kemp - From the Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599-7260 and
Joyce T Reardon - From the Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599-7260 and
Aziz Sancar - From the Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599-7260 and
Publication Details: The Journal of biological chemistry, Vol.288(29), pp.20918-20926
Academic Unit: UNKNOWN
Publisher: United States
Grant note: GM32833 / NIGMS NIH HHS P30 ES010126 / NIEHS NIH HHS K99 ES022640 / NIEHS NIH HHS R01 GM032833 / NIGMS NIH HHS
Identifiers: 99900546960401842
Language: English
Resource Type: Journal article