DNA Repair 2 (2003) 325–336
DNA bending by the human damage recognition
, Kaoru Sugasawa
, Yuichiro Shimizu
, Fumio Hanaoka
, Miodrag Djurica
, Jan H.J. Hoeijmakers
, Claire Wyman
Department of Cell Biology and Genetics, Erasmus MC, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands
Cellular Physiology Laboratory, RIKEN (The Institute of Physical and Chemical Research),
2-1 Hirosawa, Wako, Saitama 351-0198, Japan
CREST, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan
Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamada-oka, Suita, Osaka 565-0871, Japan
KPN Research, P.O. Box 421, 2260 AK Leidschendam, The Netherlands
Department of Radiation Oncology, Erasmus MC-Daniel, Rotterdam, The Netherlands
Received 12 August 2002; received in revised form 5 November 2002; accepted 11 November 2002
Genome integrity is maintained, despite constant assault on DNA, due to the action of a variety of DNA repair pathways.
Nucleotide excision repair (NER) protects the genome from the deleterious effects of UV irradiation as well as other agents
that induce chemical changes in DNA bases. The mechanistic steps required for eukaryotic NER involve the concerted action
of at least six proteins or protein complexes. The speciﬁcity to incise only the DNA strand including the damage at deﬁned
positions is determined by the coordinated assembly of active protein complexes onto damaged DNA. In order to understand
the molecular mechanism of the NER reactions and the origin of this speciﬁcity and control we analyzed the architecture
of functional NER complexes at nanometer resolution by scanning force microscopy (SFM). In the initial step of damage
recognition by XPC–HR23B we observe a protein induced change in DNA conformation. XPC–HR23B induces a bend in
DNA upon binding and this is stabilized at the site of damage. We discuss the importance of the XPC–HR23B-induced
distortion as an architectural feature that can be exploited for subsequent assembly of an active NER complex.
© 2002 Elsevier Science B.V. All rights reserved.
Keywords: Nucleotide excision repair; Damage recognition; Scanning force microscopy; XPC; DNA bending
Nucleotide excision repair (NER) eliminates of a
wide variety of lesions from DNA thereby protecting
Abbreviations: NER, nucleotide excision repair; GGR, global
genome repair; TCR, transcription coupled repair; SFM, scanning
force microscopy; CFP, cyan ﬂuorescent protein
Corresponding author. Tel.: +31-10-4088337;
E-mail address: email@example.com (C. Wyman).
virtually all organisms from the severe consequences
of DNA injuries including cell death and mutagenesis.
In humans, impaired NER activity is responsible for
the sensitivity to sunlight, increased incidence of skin
cancer and neurodegeneration experienced by xero-
derma pigmentosum (XP) patients. The different genes
involved in XP, XPA through XPG, encode products
that are required for proper NER. Mechanistically the
complete NER reaction can be separated into a series
of distinct steps: (1) damage recognition, (2) damage
1568-7864/02/$ – see front matter © 2002 Elsevier Science B.V. All rights reserved.