Human nucleotide excision repair protein XPA: Extended X-ray absorption fine-structure evidence for a metal-binding domain
Nancy J. Hess
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Search for more papers by this authorGarry W. Buchko
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Search for more papers by this authorShuisong Ni
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Search for more papers by this authorBrian D. Thrall
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Search for more papers by this authorCorresponding Author
Michael A. Kennedy
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Pacific Northwest National Laboratories, Environmental Molecular Sciences Laboratory, Richland, Washington 99352Search for more papers by this authorSteven D. Conradson
Los Alamos National Laboratories, Materials Science and Technology Division, Los Alamos, New Mexico 89575
Search for more papers by this authorF. Javier Espinosa
Los Alamos National Laboratories, Materials Science and Technology Division, Los Alamos, New Mexico 89575
Search for more papers by this authorNancy J. Hess
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Search for more papers by this authorGarry W. Buchko
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Search for more papers by this authorShuisong Ni
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Search for more papers by this authorBrian D. Thrall
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Search for more papers by this authorCorresponding Author
Michael A. Kennedy
Pacific Northwest National Laboratories, Biogeochemistry Resources, Environmental Molecular Sciences Laboratory and Molecular Biosciences Department, Richland, Washington 99352
Pacific Northwest National Laboratories, Environmental Molecular Sciences Laboratory, Richland, Washington 99352Search for more papers by this authorSteven D. Conradson
Los Alamos National Laboratories, Materials Science and Technology Division, Los Alamos, New Mexico 89575
Search for more papers by this authorF. Javier Espinosa
Los Alamos National Laboratories, Materials Science and Technology Division, Los Alamos, New Mexico 89575
Search for more papers by this authorAbstract
The ubiquitous, multi-enzyme, nucleotide excision repair (NER) pathway is responsible for correcting a wide range of chemically and structurally distinct DNA lesions in the eukaryotic genome. Human XPA, a 31 kDa, zinc-associated protein, is thought to play a major NER role in the recognition of damaged DNA and the recruitment of other proteins, including RPA, ERCC1, and TFIIH, to repair the damage. Sequence analyses and genetic evidence suggest that zinc is associated with a C4-type motif, C10S-X2-C108-X17-C126-X2-C129, located in the minimal DNA binding region of XPA (M98-F219). The zinc-associated motif is essential for damaged DNA recognition. Extended X-ray absorption fine structure (EXAFS) spectra collected on the zinc associated minimal DNA-binding domain of XPA (ZnXPA-MBD) show directly, for the first time, that the zinc is coordinated to the sulfur atoms of four cysteine residues with an average Zn-S bond length of 2.34 ± 0.01 Å. XPA-MBD was also expressed in minimal medium supplemented with cobalt nitrate to yield a blue-colored protein that was primarily (>95%) cobalt associated (CoXPA-MBD). EXAFS spectra collected on CoXPA-MBD show that the cobalt is also coordinated to the sulfur atoms of four cysteine residues with an average Co-S bond length of 2.33 ± 0.02 Å.
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