Header

UZH-Logo

Maintenance Infos

Double-check probing of DNA bending and unwinding by XPA-RPA: an architectural function in DNA repair.


Missura, M; Buterin, T; Hindges, R; Hübscher, U; Kaspárková, J; Brabec, V; Naegeli, H (2001). Double-check probing of DNA bending and unwinding by XPA-RPA: an architectural function in DNA repair. The EMBO Journal, 20(23):3554-3564.

Abstract

The multiprotein factor composed of XPA and replication protein A (RPA) is an essential subunit of the mammalian nucleotide excision repair system. Although XPA-RPA has been implicated in damage recognition, its activity in the DNA repair pathway remains controversial. By replacing DNA adducts with mispaired bases or non-hybridizing analogues, we found that the weak preference of XPA and RPA for damaged substrates is entirely mediated by indirect readout of DNA helix conformations. Further screening with artificially distorted substrates revealed that XPA binds most efficiently to rigidly bent duplexes but not to single-stranded DNA. Conversely, RPA recognizes single-stranded sites but not backbone bending. Thus, the association of XPA with RPA generates a double-check sensor that detects, simultaneously, backbone and base pair distortion of DNA. The affinity of XPA for sharply bent duplexes, characteristic of architectural proteins, is not compatible with a direct function during recognition of nucleotide lesions. Instead, XPA in conjunction with RPA may constitute a regulatory factor that monitors DNA bending and unwinding to verify the damage-specific localization of repair complexes or control their correct three-dimensional assembly.

Abstract

The multiprotein factor composed of XPA and replication protein A (RPA) is an essential subunit of the mammalian nucleotide excision repair system. Although XPA-RPA has been implicated in damage recognition, its activity in the DNA repair pathway remains controversial. By replacing DNA adducts with mispaired bases or non-hybridizing analogues, we found that the weak preference of XPA and RPA for damaged substrates is entirely mediated by indirect readout of DNA helix conformations. Further screening with artificially distorted substrates revealed that XPA binds most efficiently to rigidly bent duplexes but not to single-stranded DNA. Conversely, RPA recognizes single-stranded sites but not backbone bending. Thus, the association of XPA with RPA generates a double-check sensor that detects, simultaneously, backbone and base pair distortion of DNA. The affinity of XPA for sharply bent duplexes, characteristic of architectural proteins, is not compatible with a direct function during recognition of nucleotide lesions. Instead, XPA in conjunction with RPA may constitute a regulatory factor that monitors DNA bending and unwinding to verify the damage-specific localization of repair complexes or control their correct three-dimensional assembly.

Statistics

Citations

124 citations in Web of Science®
131 citations in Scopus®
Google Scholar™

Altmetrics

Downloads

99 downloads since deposited on 11 Feb 2008
22 downloads since 12 months
Detailed statistics

Additional indexing

Item Type:Journal Article, refereed
Communities & Collections:05 Vetsuisse Faculty > Department of Molecular Mechanisms of Disease
07 Faculty of Science > Department of Molecular Mechanisms of Disease

05 Vetsuisse Faculty > Institute of Veterinary Pharmacology and Toxicology
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:2 July 2001
Deposited On:11 Feb 2008 12:18
Last Modified:21 Nov 2017 13:22
Publisher:European Molecular Biology Organization ; Nature Publishing Group
ISSN:0261-4189
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.1093/emboj/20.13.3554
Related URLs:http://www.nature.com/emboj/index.html
http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=11432842
PubMed ID:11432842

Download

Download PDF  'Double-check probing of DNA bending and unwinding by XPA-RPA: an architectural function in DNA repair.'.
Preview
Filetype: PDF
Size: 406kB
View at publisher