FANCI and FANCD2 Have Common As Well As Independent Functions During the Cellular Replication Stress Response

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2017 Oct 24

PMID 29059323

Citations 21

Authors

Elizabeth L Thompson

Jung E Yeo

Eun-A Lee

Yinan Kan

Maya Raghunandan

Constanze Wiek

Helmut Hanenberg

Orlando D Scharer

Eric A Hendrickson

Alexandra Sobeck

Affiliations

Soon will be listed here.

Abstract

Fanconi anemia (FA) is an inherited cancer predisposition syndrome characterized by cellular hypersensitivity to DNA interstrand crosslinks (ICLs). To repair these lesions, the FA proteins act in a linear hierarchy: following ICL detection on chromatin, the FA core complex monoubiquitinates and recruits the central FANCI and FANCD2 proteins that subsequently coordinate ICL removal and repair of the ensuing DNA double-stranded break by homology-dependent repair (HDR). FANCD2 also functions during the replication stress response by mediating the restart of temporarily stalled replication forks thereby suppressing the firing of new replication origins. To address if FANCI is also involved in these FANCD2-dependent mechanisms, we generated isogenic FANCI-, FANCD2- and FANCI:FANCD2 double-null cells. We show that FANCI and FANCD2 are partially independent regarding their protein stability, nuclear localization and chromatin recruitment and contribute independently to cellular proliferation. Simultaneously, FANCD2-but not FANCI-plays a major role in HDR-mediated replication restart and in suppressing new origin firing. Consistent with this observation, deficiencies in HDR-mediated DNA DSB repair can be overcome by stabilizing RAD51 filament formation in cells lacking functional FANCD2. We propose that FANCI and FANCD2 have partially non-overlapping and possibly even opposing roles during the replication stress response.

Citing Articles

Knockdown of FANCI suppresses hepatocellular carcinoma development via the PI3K/Akt/GSK-3β pathway.

Yin Z, Lu M, Fu R Heliyon. 2025; 11(4):e42731.

PMID: 40040987 PMC: 11876927. DOI: 10.1016/j.heliyon.2025.e42731.

Cell Type Specific Suppression of Hyper-Recombination by Human RAD18 Is Linked to Proliferating Cell Nuclear Antigen K164 Ubiquitination.

Rogers C, Leung W, Baxley R, Kram R, Wang L, Buytendorp J Biomolecules. 2025; 15(1.

PMID: 39858544 PMC: 11763143. DOI: 10.3390/biom15010150.

Deregulated protein homeostasis constrains fetal hematopoietic stem cell pool expansion in Fanconi anemia.

Kovuru N, Mochizuki-Kashio M, Menna T, Jeffrey G, Hong Y, Yoon Y Nat Commun. 2024; 15(1):1852.

PMID: 38424108 PMC: 10904799. DOI: 10.1038/s41467-024-46159-1.

Mitotic DNA Synthesis in Untransformed Human Cells Preserves Common Fragile Site Stability via a FANCD2-Driven Mechanism That Requires HELQ.

Traband E, Hammerlund S, Shameem M, Narayan A, Ramana S, Tella A J Mol Biol. 2023; 435(22):168294.

PMID: 37777152 PMC: 10839910. DOI: 10.1016/j.jmb.2023.168294.

FANCD2 and RAD51 recombinase directly inhibit DNA2 nuclease at stalled replication forks and FANCD2 acts as a novel RAD51 mediator in strand exchange to promote genome stability.

Liu W, Polaczek P, Roubal I, Meng Y, Choe W, Caron M Nucleic Acids Res. 2023; 51(17):9144-9165.

PMID: 37526271 PMC: 10516637. DOI: 10.1093/nar/gkad624.

References

Klein Douwel D, Boonen R, Long D, Szypowska A, Raschle M, Walter J . XPF-ERCC1 acts in Unhooking DNA interstrand crosslinks in cooperation with FANCD2 and FANCP/SLX4. Mol Cell. 2014; 54(3):460-71. PMC: 5067070. DOI: 10.1016/j.molcel.2014.03.015. View

Kohli M, Rago C, Lengauer C, Kinzler K, Vogelstein B . Facile methods for generating human somatic cell gene knockouts using recombinant adeno-associated viruses. Nucleic Acids Res. 2004; 32(1):e3. PMC: 373311. DOI: 10.1093/nar/gnh009. View

Smogorzewska A, Matsuoka S, Vinciguerra P, McDonald 3rd E, Hurov K, Luo J . Identification of the FANCI protein, a monoubiquitinated FANCD2 paralog required for DNA repair. Cell. 2007; 129(2):289-301. PMC: 2175179. DOI: 10.1016/j.cell.2007.03.009. View

Petermann E, Orta M, Issaeva N, Schultz N, Helleday T . Hydroxyurea-stalled replication forks become progressively inactivated and require two different RAD51-mediated pathways for restart and repair. Mol Cell. 2010; 37(4):492-502. PMC: 2958316. DOI: 10.1016/j.molcel.2010.01.021. View

Kupfer G . Fanconi anemia: a signal transduction and DNA repair pathway. Yale J Biol Med. 2013; 86(4):491-7. PMC: 3848103. View

Colnaghi L, Jones M, Cotto-Rios X, Schindler D, Hanenberg H, Huang T . Patient-derived C-terminal mutation of FANCI causes protein mislocalization and reveals putative EDGE motif function in DNA repair. Blood. 2010; 117(7):2247-56. PMC: 3062332. DOI: 10.1182/blood-2010-07-295758. View

Niraj J, Caron M, Drapeau K, Berube S, Guitton-Sert L, Coulombe Y . The identification of FANCD2 DNA binding domains reveals nuclear localization sequences. Nucleic Acids Res. 2017; 45(14):8341-8357. PMC: 5737651. DOI: 10.1093/nar/gkx543. View

Chen X, Bosques L, Sung P, Kupfer G . A novel role for non-ubiquitinated FANCD2 in response to hydroxyurea-induced DNA damage. Oncogene. 2015; 35(1):22-34. PMC: 5450631. DOI: 10.1038/onc.2015.68. View

Joksic I, Vujic D, Guc-Scekic M, Leskovac A, Petrovic S, Ojani M . Dysfunctional telomeres in primary cells from Fanconi anemia FANCD2 patients. Genome Integr. 2012; 3(1):6. PMC: 3511208. DOI: 10.1186/2041-9414-3-6. View

10.

Williams S, Longerich S, Sung P, Vaziri C, Kupfer G . The E3 ubiquitin ligase RAD18 regulates ubiquitylation and chromatin loading of FANCD2 and FANCI. Blood. 2011; 117(19):5078-87. PMC: 3109534. DOI: 10.1182/blood-2010-10-311761. View

11.

Oh S, Wang Y, Zimbric J, Hendrickson E . Human LIGIV is synthetically lethal with the loss of Rad54B-dependent recombination and is required for certain chromosome fusion events induced by telomere dysfunction. Nucleic Acids Res. 2013; 41(3):1734-49. PMC: 3561972. DOI: 10.1093/nar/gks1326. View

12.

Digweed M, Rothe S, Demuth I, Scholz R, Schindler D, Stumm M . Attenuation of the formation of DNA-repair foci containing RAD51 in Fanconi anaemia. Carcinogenesis. 2002; 23(7):1121-6. DOI: 10.1093/carcin/23.7.1121. View

13.

Xu D, Guo R, Sobeck A, Bachrati C, Yang J, Enomoto T . RMI, a new OB-fold complex essential for Bloom syndrome protein to maintain genome stability. Genes Dev. 2008; 22(20):2843-55. PMC: 2569887. DOI: 10.1101/gad.1708608. View

14.

Nakanishi K, Yang Y, Pierce A, Taniguchi T, Digweed M, DAndrea A . Human Fanconi anemia monoubiquitination pathway promotes homologous DNA repair. Proc Natl Acad Sci U S A. 2005; 102(4):1110-5. PMC: 545844. DOI: 10.1073/pnas.0407796102. View

15.

Davies S, North P, Hickson I . Role for BLM in replication-fork restart and suppression of origin firing after replicative stress. Nat Struct Mol Biol. 2007; 14(7):677-9. DOI: 10.1038/nsmb1267. View

16.

Ran F, Hsu P, Wright J, Agarwala V, Scott D, Zhang F . Genome engineering using the CRISPR-Cas9 system. Nat Protoc. 2013; 8(11):2281-2308. PMC: 3969860. DOI: 10.1038/nprot.2013.143. View

17.

Castella M, Jacquemont C, Thompson E, Yeo J, Cheung R, Huang J . FANCI Regulates Recruitment of the FA Core Complex at Sites of DNA Damage Independently of FANCD2. PLoS Genet. 2015; 11(10):e1005563. PMC: 4592014. DOI: 10.1371/journal.pgen.1005563. View

18.

Schlacher K, Christ N, Siaud N, Egashira A, Wu H, Jasin M . Double-strand break repair-independent role for BRCA2 in blocking stalled replication fork degradation by MRE11. Cell. 2011; 145(4):529-42. PMC: 3261725. DOI: 10.1016/j.cell.2011.03.041. View

19.

Wang A, Kim T, Wagner J, Conti B, Lach F, Huang A . A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination. Mol Cell. 2015; 59(3):478-90. PMC: 4529964. DOI: 10.1016/j.molcel.2015.07.009. View

20.

Michl J, Zimmer J, Tarsounas M . Interplay between Fanconi anemia and homologous recombination pathways in genome integrity. EMBO J. 2016; 35(9):909-23. PMC: 4865030. DOI: 10.15252/embj.201693860. View