DNA damage repair

The Fanconi anaemia pathway: new players and new functions

Journal name:
Nature Reviews Molecular Cell Biology
Year published:
Published online


The Fanconi anaemia pathway repairs DNA interstrand crosslinks (ICLs) in the genome. Our understanding of this complex pathway is still evolving, as new components continue to be identified and new biochemical systems are used to elucidate the molecular steps of repair. The Fanconi anaemia pathway uses components of other known DNA repair processes to achieve proper repair of ICLs. Moreover, Fanconi anaemia proteins have functions in genome maintenance beyond their canonical roles of repairing ICLs. Such functions include the stabilization of replication forks and the regulation of cytokinesis. Thus, Fanconi anaemia proteins are emerging as master regulators of genomic integrity that coordinate several repair processes. Here, we summarize our current understanding of the functions of the Fanconi anaemia pathway in ICL repair, together with an overview of its connections with other repair pathways and its emerging roles in genome maintenance.

At a glance


  1. Cooperation of Fanconi anaemia, nucleotide excision repair, translesion synthesis and homologous recombination proteins in a common interstrand crosslink repair pathway.
    Figure 1: Cooperation of Fanconi anaemia, nucleotide excision repair, translesion synthesis and homologous recombination proteins in a common interstrand crosslink repair pathway.

    Stalling of replication forks on DNA interstrand crosslinks (ICLs) induces lesion recognition by the FANCM–FAAP24–MHF1–MHF2 complex (not shown) and subsequent recruitment of the Fanconi anaemia core complex. UHRF1 (ubiquitin-like with PHD and RING finger domains 1) might also be involved in lesion sensing. FANCM promotes an ATR (ataxia telangiectasia and RAD3-related) kinase-dependent checkpoint response, which in turn phosphorylates and activates multiple Fanconi anaemia proteins. A consequence of activation of the Fanconi anaemia core complex is the monoubiquitylation of the FANCD2–FANCI (FANCD2–I) heterodimer, which is promoted by the ubiquitin ligase FANCL and its partner ubiquitin-conjugating enzyme E2 T (UBE2T). ICLs in the S phase of the cell cycle impede replication fork progression, and leading strands pause 20–40 nucleotides away on either side of the ICL. Eviction of the replicative helicase CMG complex through the action of breast cancer type 1 susceptibility protein (BRCA1) allows the approach of one replication fork to within one nucleotide of the ICL. Ubiquitylated FANCD2 is directed to the ICL region, where it functions as a landing pad for the recruitment of several factors, including SLX4 and Fanconi-associated nuclease 1 (FAN1), and coordinates nucleolytic incisions that are probably mediated by ERCC4 (a structure-specific endonuclease that also functions in nucleotide excision repair) and possibly MUS81. Unhooking the DNA leaves the crosslinked nucleotide tethered to the complementary strand, which is bypassed by translesion synthesis polymerases such as REV1 or DNA polymerase ζ (REV3–REV7). Ligation restores an intact DNA duplex, which functions as a template for homologous recombination-mediated repair of the double-strand break (DSB). The DNA incisions create a DSB, which is further processed by nucleases such as CtBP-interacting protein (CtIP), MRN (MRE11–RAD50–NBS1), exonuclease 1 (EXO1) and the helicase–nuclease complex BLM–DNA2 (Bloom syndrome protein–DNA replication ATP-dependent helicase/nuclease 2) that create a single-stranded DNA (ssDNA) overhang. This ssDNA coated with replication protein A (RPA) is a substrate for RAD51-mediated strand invasion promoted by BRCA2 and subsequent homologous recombination. The USP1–UAF1 (ubiquitin carboxyl-terminal hydrolase 1– USP1-associated factor 1) complex deubiquitylates the FANCD2–I heterodimer and completes repair.

  2. The Fanconi anaemia pathway has a key role in stabilizing stalled replication forks.
    Figure 2: The Fanconi anaemia pathway has a key role in stabilizing stalled replication forks.

    In the S phase of the cell cycle, the ubiquitylated FANCD2–FANCI heterodimer (FANCD2–I-Ub) and the core Fanconi anaemia proteins FANCA, FANCC, FANCJ and FANCM (FANCA–C–J–M), together with the Fanconi anaemia-associated protein BOD1L (biorientation of chromosomes in cell division protein 1-like), localize to replication forks and bind the nascent strand (single-stranded DNA), thus protecting it from double-strand break (DSB) repair protein MRE11- and/or DNA2 (DNA replication ATP-dependent helicase/nuclease 2)-mediated nucleolytic degradation. The downstream Fanconi anaemia proteins breast cancer type 1 susceptibility protein (BRCA1), BRCA2 and RAD51 also function in fork protection. This function is crucial to preserve genome stability and is thought to depend on the ATR (ataxia telangiectasia and RAD3-related) and ATM (ataxia telangiectasia mutated) kinases. Fanconi anaemia pathway-mediated fork stability includes protection from nucleolytic degradation, and suppression of new origin firing and mitotic entry. Conversely, fork destabilization induces fork collapse, DSB formation, increased genomic instability and cell death. The mechanisms by which Fanconi anaemia proteins are recruited to stalled forks are unclear.

  3. Crosstalk between the Fanconi anaemia pathway and other repair processes.
    Figure 3: Crosstalk between the Fanconi anaemia pathway and other repair processes.

    The FANCD2–FANCI (FANCD2-I) complex is thought to antagonize non-homologous end joining (NHEJ) through an interaction with Ku proteins. This crosstalk might also involve other Fanconi anaemia proteins. The Fanconi anaemia proteins, particularly FANCD2, maintain genomic stability by ensuring the proper segregation of chromosomes during mitosis. Moreover, the Fanconi anaemia pathway interfaces with several other repair processes. First, there is increasing evidence that the Fanconi anaemia proteins, including ubiquitylated FANCD2 (FANCD2-Ub), have a crucial role in protecting replication forks from nucleolytic degradation. This fork protection probably occurs either upstream of or contemporaneously with other repair mechanisms. Second, FANCD2 recruits the nucleotide excision repair factor ERCC4 via SLX4, and XPA (xeroderma pigmentosum group A-complementing protein) and XPC may promote recruitment of the Fanconi anaemia core complex (not shown). Moreover, FANCD2-I is activated by the translesion synthesis factor RAD18 through the monoubiquitylation of proliferating cell nuclear antigen (PCNA), and Fanconi anaemia proteins facilitate the recruitment of translesion synthesis polymerases to interstrand crosslink (ICL) lesions through the interaction of Fanconi anaemia-associated protein 20 (FAAP20) with DNA repair protein REV1. The homologous recombination proteins breast cancer type 1 susceptibility protein (BRCA1), BRCA2, FANCJ, partner and localizer of BRCA2 (PALB2) and RAD51 are also Fanconi anaemia proteins, and FANCD2 is thought to function in alternative end joining, possibly by favouring the recruitment of the alternative end-joining factor DNA polymerase θ (Pol θ) to sites of DNA damage. For the sake of simplicity, the crosstalk mechanisms are shown at the replication fork; however, these could also occur in other contexts or cell cycle phases.


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  1. Department of Radiation Oncology and Center for DNA Damage and Repair, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02215, USA.

    • Raphael Ceccaldi,
    • Prabha Sarangi &
    • Alan D. D'Andrea

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The authors declare no competing interests.

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  • Raphael Ceccaldi

    Raphael Ceccaldi received dual degrees in Pharmacy (Pharm.D.) and Haematology (Ph.D.) from the University of Paris V and Paris Diderot-Paris 7, France, and is currently an instructor in the Department of Radiation Oncology at the Dana-Farber Cancer Institute, Boston, Massachusetts, USA. Raphael Ceccaldi's main research focus is to understand the molecular mechanisms underlying crosstalk between DNA repair processes.

  • Prabha Sarangi

    Prabha Sarangi received her undergraduate degree in Biotechnology from the Indian Institute of Technology, Madras, India, and her Ph.D. from Cornell University, New York, USA. She is now a postdoctoral fellow in the Department of Radiation Oncology at the Dana-Farber Cancer Institute, Boston, Massachusetts, USA.

  • Alan D. D'Andrea

    A graduate of Harvard College, Alan D. D'Andrea received his M.D. from Harvard Medical School, Boston, Massachusetts, USA, in 1983. He completed his residency in Pediatrics at the Children's Hospital of Philadelphia, USA, and a fellowship in pediatric haematology–oncology at the Dana-Farber Cancer Institute (DFCI) and Children's Hospital, Boston. He also completed a research fellowship at the Whitehead Institute of Biomedical Research at the Massachusetts Institute of Technology, where he cloned the receptor for erythropoietin while working in the laboratory of Harvey Lodish. Alan D'Andrea joined the staff at DFCI in 1990. His research is focused on the molecular cause of leukaemia. He also investigates the pathogenesis of Fanconi anaemia, a human genetic disease characterized by bone marrow failure and acute myeloid leukaemia in children. He is currently the Fuller-American Cancer Society Professor of Radiation Oncology at Harvard Medical School and the Director of the Center for DNA Damage and Repair at DFCI. Alan D. D'Andrea's homepage.

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