DNA Interstrand Crosslink Repair


Crosslinking agents such as psoralens, nitrogen mustards or cisplatin are bifunctionally acting chemicals that generate some adducts as covalent linkages between complementary deoxyribonucleic acid strands. Since many agents are of importance in genetic toxicology and cancer therapy, repair of interstrand crosslinks has been studied extensively in bacteria and in lower and higher eukaryotes. The main repair pathway in Escherichia coli involves the sequential action of nucleotide excision repair and recombinational repair. In eukaryotes, nucleotide excision repair, translesion synthesis and homologous recombination all play important roles in crosslink repair. Instances where crosslink unhooking is performed by a glycosylase have also been observed. The relative pathway contribution depends on cell cycle position and agent used. Analysis of mammalian crosslink response proteins that are also involved in the hereditary syndrome Fanconi anaemia resulted in a model of crosslink repair within the context of converging replicons arrested by an interstrand crosslink.

Key Concepts

  • Certain chemicals with two or more correctly spaced reactive groups can covalently link opposing DNA strands.
  • Several repair or tolerance pathways such as nucleotide excision repair, homologous recombination, translesion synthesis and even DNA glycosylases can work together to overcome such complex damage.
  • Cell cycle stage may determine the choice of repair pathway or pathway combinations.
  • A heritable human cancer‐prone syndrome with diverse phenotypes (Fanconi anaemia) has been linked to defects in crosslink repair.
  • By integrating in vitro studies and the analysis of Fanconi proteins, a model for mammalian cells places replication‐coupled crosslink repair at the convergence point of stalled replication forks.
  • This model includes a sequence of unhooking of the interstrand crosslink by structure‐specific endonucleases, filling of the created single‐stranded gap by translesion synthesis and repair of the double‐strand break in the sister chromatid by one of several pathways, including homologous recombination.

Keywords: DNA repair; crosslinks; excision; bifunctional alkylation; recombination; translesion synthesis; Fanconi anaemia

Figure 1. Examples of crosslinking agents and their primary interstrand crosslink adduct. (a) Nitrogen mustard, (b) mitomycin C, (c) cisplatin, (d) methoxypsoralen, (e) nitrosourea, (f) diepoxybutane, (g) aldehyde and (h) nitric oxide. From Lopez‐Martinez D, Liang CC and Cohn MA (2016) Cellular response to DNA interstrand crosslinks: the Fanconi anemia pathway. Cellular and Molecular Life Sciences 73: 3097–3114, Licensed under CC BY.
Figure 2. Repair of an ICL in E. coli. (a) In the main pathway, first, a single‐stranded DNA fragment is removed by NER by cutting 3′ and 5′ of the crosslink. It stays attached to the complementary strand through the crosslink. A gap is formed by the exonuclease activity of Pol I. The single‐stranded gap is filled by RecA‐dependent strand invasion as the initial step of HR, and a second round of NER releases a partially double‐stranded fragment containing the crosslink. (b) The minor pathway found in stationary cells also starts with crosslink excision. Then, however, DNA polymerase II (β) performs translesion synthesis and a second round of NER releases the same product as in (a). From Dronkert ML and Kanaar R Repair of DNA interstrand crosslinks. Mutation Research 486: 217–247 © Elsevier.
Figure 3. Model for replication‐coupled DNA crosslink repair, based on studies of a cell‐free system. (a) Plasmid substrate, containing a single ICL in a defined position.From Räschle M, Knipscheer P, Enoiu M et al. (2008) Mechanism of replication‐coupled DNA interstrand crosslink repair. Cell 134: 969–980. © Elsevier. (b) Electron microscopy analysis of replication of this substrate, following cell extract addition.Reproduced with permission from Räschle et al. . © Cell Press and Elsevier. (c) Scheme for crosslink processing. The assumption of converging and stalled replication forks may lead to simultaneous double‐strand breakage and unhooking of the crosslink. Compare to Figure , a similar scheme that includes candidate proteins. From Räschle M, Knipscheer P, Enoiu M et al. (2008) Mechanism of replication‐coupled DNA interstrand crosslink repair. Cell 134: 969–980. © Elsevier.
Figure 4. Model for replication‐coupled DNA crosslink repair, incorporating studies on FA proteins. (a,b) Removal of the replicative CMG helicase following its ubiquitination by TRAIP sets the stage for ICL recognition by the FANCM‐containing anchoring complex. Following stabilisation of the stalled replication fork, the FA core complex joins and carries out the monoubiquitination of FANCI/FANCD2 heterodimer (ID2). (c) Through binding of the UBZ4 domain‐containing scaffold protein, SLX4 brings in structure‐specific endonucleases (ERCC1–XPF is shown) that may cut in sequential or redundant fashion to unhook the crosslink. This may also include FAN1. (d) Next, TLS is carried out by REV1/Polζ (mainly). The unhooked crosslink may later undergo complete removal by NER. (e) The double‐strand break in the sister chromatid will undergo end resection and repair by HR, microhomology‐mediated or nonhomologous end joining (the details of these processes are not subject of this review). From Liu W, Palovcak A, Li F, Zafar A, Yuan F and Zhang Y (2020) Fanconi anemia pathway as a prospective target for cancer intervention. Cell & Bioscience 10: 39. Licensed under CC BY 4.0.


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Siede, Wolfram(Dec 2020) DNA Interstrand Crosslink Repair. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0029226]