Mutations in fa proteins give rise to fanconi anemia, a cancer predisposition syndrome. Mechanism of replicationcoupled dna interstrand crosslink. Mechanisms of interstrand dna crosslink repair and human disorders. Replication fork reversal during dna interstrand crosslink. Many cancer chemotherapeutic agents form dna interstrand crosslinks icls, extremely cytotoxic lesions that form covalent bonds between two opposing dna strands, blocking dna replication and transcription. Pdf dna interstrand crosslink repair and cancer researchgate. The use of dna interstrand crosslinking agents as a. Dna interstrand crosslink repair siede major reference. Dna interstrand crosslinks icls are toxic dna lesions whose repair occurs in the s phase of metazoans via an unknown mechanism. This crosslink can occur within the same strand intrastrand or between opposite strands of doublestranded dna interstrand. Therefore, drugs inducing icls are effective against rapidly growing cancer cells. In metazoans, a major pathway of icl repair is coupled to dna replication, and it requires the fanconi anemia pathway. Recent studies have revealed the mechanisms and the. Mechanism of rad51dependent dna interstrand crosslink repair.
We show that a functionally uncharacterized protein, hsf2bp, is involved in a novel, direct and highly evolutionarily conserved interaction with brca2. Dna interstrand crosslinks icls are a particularly serious form of dna damage that can be formed by anti cancer alkylating agentsorendogenouscompounds,includingaldehydesgaraycoechea et al. Involvement of translesion synthesis dna polymerases in dna. Dna damage response and cancer therapeutics through the lens of the fanconi anemia dna repair pathway sonali bhattacharjee and saikat nandi abstract fanconi anemia fa is a rare, inherited genomic instability disorder, caused by mutations in genes involved in the repair of interstrand dna crosslinks icls. Interstrand crosslink repair as a target for hdac inhibition. Indirect evidence suggests that in metazoans, the principal. Crosslinks can be formed on the same strand of dna intrastrand, between the two complementary strands of dna interstrand, or between a base on dna and a reactive group on a protein dnaprotein.
Request pdf on jan 22, 20, taraswi banerjee and others published fanconi anemia, interstrand crosslink repair and cancer find, read and cite all the research you need on researchgate. Review open access mechanisms of interstrand dna crosslink repair and human disorders satoru hashimoto1, hirofumi anai2 and katsuhiro hanada2 abstract interstrand dna crosslinks icls are the link between watsoncrick strands of dnas with the covalent bond and. This enables identification of the sequential steps of repair mechanisms. Although it is generally known that mitochondria are defective in dna damage processing, little is known about the dna repair pathways and mechanisms that exist in these vital organelles. Repair of icls involves multiple dna repair pathways. Repair of dna interstrand crosslinks produced by cancer. A new varietal of dna interstrand crosslink repair cell.
Cisplatin is a widely used anticancer drug that acts by binding dna and causing the formation of intrastrand and interstrand icl crosslinks, but the precise downstream effects of the latter damage are not well understood. Dna interstrand crosslinks icls are formed by bifunctional agents and covalently link two strands of a dna duplex. Dna interstrand crosslinks induced by acetaldehyde are repaired by both the fanconi anaemia pathway and by a second, excisionindependent repair mechanism. Adpribosylation by adpribosyltransferases arts has a wellestablished role in dna strand break repair by promoting enrichment of repair factors at damage sites through adpribose interaction domains. Dec 18, 2009 fanconi anemia is a human cancer predisposition syndrome caused by mutations in fanc genes. This pathway includes nucleotide excision repair and homologous recombination, which is error. Dna interstrand crosslink repair in mammalian cells. However, tumors engage a complicated enzymatic machinery to repair and. Dna interstrand crosslinks icls prevent strand separation during dna replication and transcription and therefore are extremely cytotoxic. Dna interstrand crosslinks icls are among the most toxic dna damages. In genetics, crosslinking of dna occurs when various exogenous or endogenous agents react with two nucleotides of dna, forming a covalent linkage between them. Here, we describe a cellfree system based on xenopus egg extracts that supports icl repair.
The fanconi anemia pathway promotes replicationdependent dna. In vertebrate cells, dna interstrand crosslink icl repair is coupled to dna replication and involves structurespecific endonucleases, translesion dna polymerases, recombinases, and numerous proteins mutated in the human disease fanconi anemia fa. Hsf2bp negatively regulates homologous recombination in dna. Pdf the colibactin genotoxin generates dna interstrand.
Interstrand crosslinks, which link the two complementary strands of dna together, comprise less than 5% of the total dna lesions but are highly cytotoxic and difficult to repair. Dna interstrand crosslinkers, a chemically diverse group of compounds which also induce alkylation of bases and dna intrastrand crosslinks, are extensively utilized for cancer therapy. Introduction dna interstrand crosslinks icls are covalent linkages between both strands of the double helix and a. Dna interstrand crosslink repair and cancer andrew j. Interstrand crosslinks icls are a type of dna damage that is induced by various chemotherapeutics, such as cisplatin. Intrastrand crosslinks are repaired by nucleotide excision repair ner, whereas icls are removed by icl repair, a process less well understood than ner 6. West abstract interstrand crosslinks icls are highly toxic dna lesions that prevent transcription and. We have recently shown enhanced repair of dna interstrand crosslinks in ovarian cancer cells from patients following treatment with platinumbased chemotherapy 9. Rpa has previously been shown to play a role in recognizing and binding damaged dna during nucleotide excision repair ner.
Although hsf2bp was previously described as testisspecific, we find it is. Repair of dna interstrand crosslinks sciencedirect. In contrast, the percentage repair unhooking of dna interstrand crosslinks was much greater in the group of treated patients. Brca1 functions independently of homologous recombination in dna interstrand crosslink repair. Molecular targets and clinical applications, second edition provides a comprehensive and timely reference that focuses on the translational and clinical use of dna repair as a target area for the development of diagnostic biomarkers and the enhancement of cancer treatment. Interstrand dna crosslinks icls are the link between watsoncrick strands of dnas with the covalent bond and prevent separation of dna strands. Dna interstrand crosslink repair during g1 involves nucleotide excision repair and dna polymerase zeta. Koichi sato, inger brandsma, sari e van rossumfikkert, nicole verkaik, anneke b oostra, josephine c dorsman, dik c van gent, puck knipscheer, roland kanaar, alex n zelensky, hsf2bp negatively regulates homologous recombination in dna interstrand crosslink repair, nucleic acids research, volume 48, issue 5, 18 march 2020, pages 24422456. Failure to repair endogenous icls in s phase is thought to underlie the bone marrow failure syndrome fanconi anemia, and upregulation of icl repair is one. Pdf interstrand crosslinks icls are highly toxic dna lesions that prevent transcription and replication by inhibiting dna strand separation. Although neil3 has been shown to unhook interstrand crosslinks icl in xenopus extracts, how neil3 participants in icl repair in human cells and its corporation with the canonical fanconi anemia fabrca pathway remain unclear.
Aug 26, 2008 replication protein a rpa is a heterotrimeric protein that is required for dna replication and most dna repair pathways. Crosslinking of dna an overview sciencedirect topics. Dandrea, in the molecular basis of cancer fourth edition, 2015. By covalently linking both dna strands, icls prevent dna melting, transcription, and replication. The neil3 dna glycosylase is a base excision repair enzyme that excises bulky base lesions from dna. A clinical rad51 mutation disrupts dna interstrand crosslink. Dna interstrand crosslinks, with emphasis on repair, cell. Repair of dna interstrand crosslinks icls predominantly involves the. In vertebrates, icls repair is coupled to replication and involves structurespeci. Agents that induce icls were one of the earliest, and are still the most widely used, forms of chemotherapeutic drug.
To investigate the possibility of measuring the genespecific dna damage after therapeutic exposure to nitrogen mustards and to examine its relationship with the clinical response. Introduction dna interstrand crosslinks icls impede dna replication and transcription and are therefore extremely cytotoxic. Dna cross linking an overview sciencedirect topics. The common feature of these diverse lesions is that two nucleotides on opposite strands are covalently joined. The role of adpribosylation in regulating dna interstrand. A dominant mutation in human rad51 reveals its function in. During dna replication of a plasmid containing a sitespecific icl, two replication forks converge on the crosslink. The ability of cells to repair dna icls is a critical determinant of sensitivity, and recent clinical studies. Certain lesions that are removed by base excision repair are efficiently removed in mitochondria, whereas some bulky lesions that are removed by nucleotide excision repair are not repaired in these organelles.
Dna interstrand crosslinks icls are toxic dna lesions whose repair in s phase of eukaryotic cells is incompletely understood. Only recently, however, have we begun to understand how cells repair these lesions. Journal dna interstrand crosslink repair during g1 involves. Nondenaturable interstrand crosslinks in dna exposed to pks e. At 24 h in the 36 newly diagnosed patient tumour samples, only one gave 50% repair and 23 gave repair. Moreover, icls can induce mutations and rearrangements of dna, possibly resulting in uncontrolled cell growth and tumor formation. The proteins involved in icl repair, in general, are required for resistance to icls and represent potential therapeutic targets. So far, nucleotide excision repair ner, structurespecific endonucleases, translesion dna synthesis tls, homologous recombination hr, and factors. Fanconi anemia, interstrand crosslink repair and cancer. Mechanisms of interstrand dna crosslink repair and human. However, cellular responses triggered by icls can cause resistance in tumor cells, limiting the efficacy of such treatment. However, tumors engage a complicated enzymatic machinery to repair and survive icls. Mechanisms of repair of interstrand crosslinks in dna. In contrast, interstrand crosslinks icls between the two dna strands are minor lesions, accounting for less than 5% of all cisplatin lesions 5.
However, brca1,53bp1 doubledeficient cells are hypersensitive to dna interstrand crosslinks icls, indicating that brca1 has an additional role in dna crosslink repair that is distinct from hr. In xenopus egg extracts, icl repair is initiated when two replication forks converge on the lesion. Understanding the cellular response to dna damage induced by these agents is critical for more effective utilization of these compounds and for the. A central event in the activation of the fanconi anemia pathway is the monoubiquitylation of the fancifancd2 complex, but how this complex confers icl. Mechanism of replicationcoupled dna interstrand crosslink repair. Dna interstrand crosslink repair and cancer nature. So far, nucleotide excision repair ner, structurespecific endonucleases, translesion dna synthesis tls, homologous recombination hr, and factors responsible.
Cooperation of the neil3 and fanconi anemiabrca pathways in. Novel crosslinking agents, and targeting strategies involving dna crosslinking agents, continue to be developed. Dna crosslinking damage and cancer a tale of friend and foe. Enhanced repair of dna interstrand crosslinking in ovarian. Molecular targets and clinical applications, second edition provides a comprehensive and timely reference that focuses on the translational and clinical use of dna repair as a target area for the development of diagnostic biomarkers and the enhancement of cancer treatment experts on dna repair proteins from all areas of cancer biology research take. Although hsf2bp was previously described as testisspecific, we find it is expressed in. Icl repair that involves a reversed fork intermediate. Defects in dna interstrand crosslink icl repair have detrimental consequences, including stem cell failure and tumorigenesis. The disorder is characterized by genomic instability and cellular hypersensitivity to chemicals that generate dna interstrand crosslinks icls. Checkpoint signaling from a single dna interstrand crosslink. Mutational inactivation of fa genes leads to developmental defects, cancer predisposition, bone marrow failure, and hypersensitivity to crosslinking agents. The proteins involved in icl repair, in general, are required for resistance to icls and. Here, we exploit the simple eukaryote dictyostelium to uncover a role for adpribosylation in regulating dna interstrand crosslink repair and redundancy of this pathway with nonhomologous end. The kinetics of genespecific monoadducts and interstrand crosslink formation repair were measured in the p53 and n ras genes.
Pmc free article this paper describes an important icl repair mechanism that is active in g1 phase of the cell cycle. The creation of localized damage in subnuclear domains of cells is the in vivo equivalent of sitespecific lesions. Genespecific formation and repair of dna monoadducts and. Function in dna interstrand crosslink repair independent of homologous recombination graphical abstract highlights d a dominantnegative mutation in rad51 is identi. Iclinducing agents are widely used in cancer chemotherapy, as exemplified by cisplatin and nitrogen mustard. Journal dna interstrand crosslink repair during g1.
Dna interstrand crosslinks icls are lesions caused by a variety of endogenous metabolites, environmental exposures, and cancer chemotherapeutic agents that have two reactive groups. The main repair pathway in escherichia coli involves the sequential action of nucleotide excision repair ner and recombinational repair. Strategies for dna interstrand crosslink repair tufts university. Oct 01, 2015 the fanconi anemia fa pathway orchestrates dna interstrand crosslink icl repair, a multistep process that involves dna incision, translesion synthesis, and homologous recombination hr. A new varietal of dna interstrand crosslink repair. Fanconi anemia proteins, dna interstrand crosslink repair. Initiation of dna interstrand crosslink repair in humans. For the bifunctional alkylating drugs, it now seems clear that the interstrand crosslink icl is the critical lesion. Fanconi anemia, dna interstrand crosslinks, homologous recombination, dna repair, dna damage responses, chemotherapy, bifunctional alkylating agents, platinum compounds abstract. Since the icl lesion affects both strands of the dna, the icl repair is not simple.
Dna interstrand crosslinks icls covalently connect complementary dna strands. Despite the clinical success of crosslinking agents in antitumor. Kim, department of pharmacology, medical research center for gene regulation, chonnam national university. Dna interstrand crosslink repair in mammalian cells mccabe. During dna replication of a plasmid containing a sitespeci.
Interstrand crosslinks icls are highly toxic dna lesions that prevent transcription and replication by inhibiting dna strand separation. Dna interstrand crosslinks icls represent a grave threat to genomic and. The fanconi anemia pathway promotes replicationdependent. Advances in understanding the complex mechanisms of dna. Dna interstrand crosslink repair during g1 involves nucleotide excision repair and dna polymerase f sovan sarkar1, adelina a davies2, helle d ulrich2 and peter j mchugh1, 1cancer research uk laboratories, weatherall institute of molecular. Consequently, dna replication and transcription are hampered, dna damage responses ddr are initiated, and cell death is triggered.
Pdf fanconi anemia proteins, dna interstrand crosslink. In this study, we investigated the influence of cisplatin icls on synthetic nucleosomes that were platinated in a sitespecific manner in vitro and on gene transcription in. Dna interstrand crosslinks icls are the most toxic lesions induced by chemotherapeutic agents such as mitomycin c and cisplatin. Dna interstrand crosslinks, with emphasis on repair, cell signaling, and therapeutic implications. Disturbed this icl repair orchestration in s phase causes genome instability resulting a cancer prone disease, fanconi anemia. Since many of these 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. Cells deficient in icl repair are hypersensitive to a variety of bifunctional alkylating agents and exhibit excessive genomic instability. The tumor suppressor brca2 is essential for homologous recombination, replication fork stability and dna interstrand crosslink icl repair in vertebrates. Seidman, dna interstrand crosslink repair in mammalian cells. Dual incisions then create a dna doublestrand break dsb in one sister chromatid, whereas lesion bypass restores the other sister. Drugs that produce dna interstrand crosslinks icls, between the two complementary strands of the double helix, have an important role in chemotherapy regimens for cancer. Dna interstrand crosslink repair during g1 involves nucleotide excision repair and dna polymerase f sovan sarkar1, adelina a davies2, helle d ulrich2 and peter j mchugh1, 1cancer research uk laboratories, weatherall institute of molecular medicine, university of. Local damage has been extremely useful for studying the recruitment of dna repair proteins to sites of dna damage volker et al. Fanconi anemia is a human cancer predisposition syndrome caused by mutations in fanc genes.
Interstrand crosslinks icls represent a major challenge for dna replication and transcription by preventing dna strand separation. Distinct cellular phenotype linked to defective dna interstrand. Dna damage response and cancer therapeutics through the lens. Interstrand dna crosslinks icls make up a particular subtype of dna lesions, and these lesions have an especially potent biological effect. Figure 1 interstrand crosslink repair pathway in prokaryotes. They can be formed by endogenous sources such as products of lipid metabolism and abasic sites, as well as by a number of antitumor agents such as nitrogen mustards, cisplatin and mitomycin c. Parameswary a muniandy, jia liu, alokes majumdar, suting liu and michael m. Dna interstrand crosslinkers, a chemically diverse group of compounds which also induce alkylation of bases and dna intrastrand crosslinks, are extensively. Sequential use of three dna repair pathways to repair dna crosslinks. Rpa has also been suggested to play a role in psoralen dna interstrand crosslink icl repair, but a clear biochemical activity has yet to be.
Inhibition of carboplatininduced dna interstrand crosslink. Studies on icl signaling and repair have been limited, because these drugs generate additional dna lesions that trigger checkpoint signaling. Dna interstrand crosslinks icls can be induced by multiple crosslinking agents and are a severe form of dna damage that prevents strand separation during dna replication and transcription. Covalent dna protein crosslinks dpcs are induced by various compounds, which include widely used anticancer drugs, and are highly cytotoxic. May 01, 2016 interstrand dna crosslinks icls are the link between watsoncrick strands of dnas with the covalent bond and prevent separation of dna strands. Walter1, 1harvard medical school, department of biological chemistry and molecular pharmacology, boston, ma 02115, usa 2institute of molecular cancer. The role of usp 1 autocleavage in dna dna interstrand crosslink repair mira kim and jung min kim department of pharmacology, medical research center for gene regulation, chonnam national university medical school, gwangju, korea correspondence j. A central event in the activation of the fanconi anemia pathway is the monoubiquitylation of the fancifancd2 complex, but how this. Fa is characterized by genomic instability and cellular sensitivity to dna interstrand cross linking agents. Dna interstrand crosslink repair requires replication. Examining even a single type of dna lesion, such as an icl, demonstrates the incredible molecular machinery cells must utilize to repair the damage. Involvement of translesion synthesis dna polymerases in. A single icl can kill repair deficient bacteria and yeast, and about 40 icls can kill repair deficient mammalian cells. Jun 28, 2017 covalent dnaprotein crosslinks dpcs are induced by various compounds, which include widely used anticancer drugs, and are highly cytotoxic.