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PMID: 17154534 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Biochemical and structural domain analysis of xeroderma pigmentosum complementation group C protein.

Biochemistry ·Vol. 45 ·No. 50 ·2006-12-19 ·Pages 14965-79

Bunick CG, Miller MR, Fuller BE, Fanning E, Chazin WJ

Abstract

XPC is a 940-residue multidomain protein critical for the sensing of aberrant DNA and initiation of global genome nucleotide excision repair. The C-terminal portion of XPC (residues 492-940; XPC-C) has critical interactions with DNA, RAD23B, CETN2, and TFIIH, whereas functional roles have not yet been assigned to the N-terminal portion (residues 1-491; XPC-N). In order to analyze the molecular basis for XPC function and mutational defects associated with xeroderma pigmentosum (XP) disease, a series of stable bacterially expressed N- and C-terminal fragments were designed on the basis of sequence analysis and produced for biochemical characterization. Limited proteolysis experiments combined with mass spectrometry revealed that the full XPC-C is stable but XPC-N is not. However, a previously unrecognized folded helical structural domain was found within XPC-N, XPC(156-325). Pull-down and protease protection assays demonstrated that XPC(156-325) physically interacts with the DNA repair factor XPA, establishing the first functional role for XPC-N. XPC-C exhibits binding characteristics of the full-length protein, including stimulation of DNA binding by physical interaction with RAD23B and CETN2. Analysis of an XPC missense mutation (Trp690Ser) found in certain patients with XP disease revealed that this mutation is associated with a diminished ability to bind DNA. Evidence of contributions to protein interactions from regions in both XPC-N and XPC-C along with recently recognized homologies to yeast PNGase prompted construction of a structural model of a folded XPC core. This model offers key insights into how domains from the two portions of the protein may cooperate in generating specific XPC functions.

MeSH Terms
Binding Sites/genetics DNA/chemistry,genetics,metabolism DNA-Binding Proteins/chemistry,genetics,metabolism Mutation, Missense Protein Binding/genetics Protein Structure, Tertiary/genetics Transcription Factors/chemistry,genetics,metabolism Xeroderma Pigmentosum/chemistry,genetics,metabolism
Chemicals
DNA-Binding Proteins Transcription Factors XPC protein, human DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Bunick Christopher G
Department of Biochemistry, Center for Structural Biology, Vanderbilt University, Nashville, Tennessee 37232-8725, USA.
Miller Michael R
Fuller Brian E
Fanning Ellen
Chazin Walter J
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Article Info
Journal
Biochemistry
Abbr.
Biochemistry
ISSN
0006-2960
DOI
10.1021/bi061370o
Published
2006-12-19
Pages
14965-79
Language
English
Region
United States
NLM ID
0370623
PMCID
PMC2579963
Subset
IM
Grants
NIGMS NIH HHS · R01 GM040120 · United States
NIGMS NIH HHS · R01 GM40120 · United States
NIEHS NIH HHS · P30 ES000267 · United States
NCI NIH HHS · P30 CA068485-06 · United States
NIGMS NIH HHS · R01 GM040120-18 · United States
NIGMS NIH HHS · R01 GM065484 · United States
NIGMS NIH HHS · T32 GM07347 · United States
NIGMS NIH HHS · R01 GM052948-11 · United States
NIGMS NIH HHS · R01 GM040120-17 · United States
NIGMS NIH HHS · R01 GM065484-03 · United States
NIGMS NIH HHS · T32 GM007347-25 · United States
NIGMS NIH HHS · R01 GM065484-04 · United States
NIGMS NIH HHS · T32 GM007347 · United States
NIEHS NIH HHS · P30 ES000267-38 · United States
NCI NIH HHS · P30 CA 68485 · United States
NCI NIH HHS · P30 CA068485 · United States
NIEHS NIH HHS · P50 ES00267 · United States
NIGMS NIH HHS · R01 GM052948-12 · United States
NIEHS NIH HHS · P30 ES000267-39 · United States
NIGMS NIH HHS · R01 GM52948 · United States
NIGMS NIH HHS · R01 GM065484-02 · United States
NIGMS NIH HHS · R01 GM052948 · United States
NIGMS NIH HHS · R01 GM65484 · United States
External Links
PubMed source DOI: 10.1021/bi061370o PMC Full Text View journal details
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