한빛사논문
- 조회1,941
Abstract
Muwen Kong1,3, Lili Liu1,3, Xuejing Chen4, Katherine I. Driscoll5, Peng Mao6, Stefanie Böhm2,3, Neil M. Kad7, Simon C. Watkins8, Kara A. Bernstein2,3, John J. Wyrick6, Jung-Hyun Min4,*, Bennett Van Houten1,3,9,*
1 Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA
2 Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA
3 University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA
4 Department of Chemistry, University of Illinois at Chicago, Chicago, IL 60607, USA
5 Department of Physics and Astronomy, University of South Carolina, Columbia, SC 29208, USA
6 School of Molecular Biosciences, Washington State University, Pullman, WA 99164, USA
7 School of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, UK
8 Center for Biologic Imaging, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA
9 Lead Contact
*Correspondence : Jung-Hyun Min, Bennett Van Houten
Summary
Nucleotide excision repair (NER) is an evolutionarily conserved mechanism that processes helix-destabilizing and/or -distorting DNA lesions, such as UV-induced photoproducts. Here, we investigate the dynamic protein-DNA interactions during the damage recognition step using single-molecule fluorescence microscopy. Quantum dot-labeled Rad4-Rad23 (yeast XPC-RAD23B ortholog) forms non-motile complexes or conducts a one-dimensional search via either random diffusion or constrained motion. Atomic force microcopy analysis of Rad4 with the β-hairpin domain 3 (BHD3) deleted reveals that this motif is non-essential for damage-specific binding and DNA bending. Furthermore, we find that deletion of seven residues in the tip of β-hairpin in BHD3 increases Rad4-Rad23 constrained motion at the expense of stable binding at sites of DNA lesions, without diminishing cellular UV resistance or photoproduct repair in vivo. These results suggest a distinct intermediate in the damage recognition process during NER, allowing dynamic DNA damage detection at a distance.
Keywords: Rad4, Rad23, XPC, nucleotide excision repair, xeroderma pigmentosum, single particle tracking, dynamic DNA damage recognition, DNA tightrope assay, quantum dots
논문정보
- Research article
- 2016년 10월 (BRIC 등록일 2016-10-13)
- 국외 연구진
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