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Human nucleotide excision repair protein XPA: NMR spectroscopic studies of an XPA fragment containing the ERCC1-binding region and the minimal DNA-binding domain (M59-F219).

Publication ,  Journal Article
Buchko, GW; Isern, NG; Spicer, LD; Kennedy, MA
Published in: Mutat Res
June 5, 2001

XPA is a central protein component of nucleotide excision repair (NER), a ubiquitous, multi-component cellular pathway responsible for the removal and repair of many structurally distinct DNA lesions from the eukaryotic genome. The solution structure of the minimal DNA-binding domain of XPA (XPA-MBD: M98-F219) has recently been determined and chemical shift mapping experiments with 15N-labeled XPA-MBD show that XPA binds DNA along a basic surface located in the C-terminal loop-rich subdomain. Here, XPA-DNA interactions are further characterized using an XPA fragment containing the minimal DNA-binding domain plus the ERCC1-binding region (XPA-EM: M59-F219). The 15N/1H HSQC spectrum of XPA-EM closely maps onto the 15N/1H HSQC spectrum of XPA-MBD, suggesting the DNA-binding domain is intact in the larger XPA fragment. Such a conclusion is corroborated by chemical shift mapping experiments of XPA-EM with a single strand DNA oligomer, dCCAATAACC (d9), that show the same set of 15N/1H HSQC cross peaks are effected by the addition of DNA. However, relative to DNA-free XPA-MBD, the 15N/1H HSQC cross peaks of many of the basic residues in the loop-rich subdomain of DNA-free XPA-EM are less intense, or gone altogether, suggesting the acidic ERRC1-binding region of XPA-EM may associate transiently with the basic DNA-binding surface. While the DNA-binding domain in XPA-EM is structured and functional, 15N-edited NOESY spectra of XPA-EM indicate that the acidic ERRC1-binding region is unstructured. If the structural features observed for XPA-EM persist in XPA, transient intramolecular association of the ERCC1-binding domain with the DNA-binding region may play a role in the sequential assembly of the NER components.

Duke Scholars

Published In

Mutat Res

DOI

ISSN

0027-5107

Publication Date

June 5, 2001

Volume

486

Issue

1

Start / End Page

1 / 10

Location

Netherlands

Related Subject Headings

  • Xeroderma Pigmentosum Group A Protein
  • Time Factors
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Protein Binding
  • Oncology & Carcinogenesis
  • Nucleic Acid Conformation
  • Models, Molecular
  • Magnetic Resonance Spectroscopy
  • Humans
 

Citation

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Buchko, G. W., Isern, N. G., Spicer, L. D., & Kennedy, M. A. (2001). Human nucleotide excision repair protein XPA: NMR spectroscopic studies of an XPA fragment containing the ERCC1-binding region and the minimal DNA-binding domain (M59-F219). Mutat Res, 486(1), 1–10. https://doi.org/10.1016/s0921-8777(01)00072-6
Buchko, G. W., N. G. Isern, L. D. Spicer, and M. A. Kennedy. “Human nucleotide excision repair protein XPA: NMR spectroscopic studies of an XPA fragment containing the ERCC1-binding region and the minimal DNA-binding domain (M59-F219).Mutat Res 486, no. 1 (June 5, 2001): 1–10. https://doi.org/10.1016/s0921-8777(01)00072-6.
Buchko, G. W., et al. “Human nucleotide excision repair protein XPA: NMR spectroscopic studies of an XPA fragment containing the ERCC1-binding region and the minimal DNA-binding domain (M59-F219).Mutat Res, vol. 486, no. 1, June 2001, pp. 1–10. Pubmed, doi:10.1016/s0921-8777(01)00072-6.
Journal cover image

Published In

Mutat Res

DOI

ISSN

0027-5107

Publication Date

June 5, 2001

Volume

486

Issue

1

Start / End Page

1 / 10

Location

Netherlands

Related Subject Headings

  • Xeroderma Pigmentosum Group A Protein
  • Time Factors
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Protein Binding
  • Oncology & Carcinogenesis
  • Nucleic Acid Conformation
  • Models, Molecular
  • Magnetic Resonance Spectroscopy
  • Humans