Biallelic mutations in the ubiquitin ligase RFWD3 cause Fanconi anemia

Abstract Background DNA-crosslinking agents like cisplatin and mitomycin C (MMC) are indispensible for the treatment of many solid malignancies. These anticancer drugs generate DNA interstrand crosslinks (ICLs) that cause cell death by blocking replication forks. Many factors counteracting ICL-induced DNA replication stress, including the Fanconi anemia (FA) pathway, are regulated by ubiquitination and, therefore, ubiquitin ligases are potential targets for the sensitization of cancer cells to crosslinking agents. In this study, we investigated the function of the CRL4 ubiquitin ligase in modulating the response of cancer cells to ICL induction. Methods The two cullin paralogs CUL4A and CUL4B, which form the CRL4 ligase scaffold, were depleted in cancer cells by small interfering RNA followed by analysis of the cellular and biochemical responses to ICLs elicited upon cisplatin or MMC treatment. Results We report that the combined depletion of CUL4A and CUL4B weakens an FA pathway-dependent S phase checkpoint response. CRL4 positively stimulates the monoubiquitination of FANCD2 required for the recruitment of XPF-ERCC1, a structure-specific endonuclease that, in turn, contributes to the display of single-stranded DNA (ssDNA) at ICLs. After CRL4 down regulation, the missing ssDNA results in reduced recruitment of RPA, thereby dampening activation of ATR and CHK1 checkpoint kinases and allowing for S phase progression despite ICL induction. Conclusion Our findings indicate that CRL4 promotes cell survival by potentiating an FA pathway-dependent ssDNA-RPA signaling platform installed at ICLs. The anticancer efficacy of crosslinking agents may, therefore, be enhanced by down regulating CRL4 activity.

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Molecular pathogenesis of Fanconi anemia: recent progress

Blood ◽

10.1182/blood-2005-10-4240 ◽

2006 ◽

Vol 107 (11) ◽

pp. 4223-4233 ◽

Cited By ~ 247

Author(s):

Toshiyasu Taniguchi ◽

Alan D. D'Andrea

Keyword(s):

Dna Repair ◽

Ubiquitin Ligase ◽

Fanconi Anemia ◽

Cancer Biology ◽

Recent Progress ◽

Cancer Susceptibility ◽

Chromosome Instability ◽

Childhood Onset ◽

Dna Crosslinking ◽

Crosslinking Agents

AbstractA rare genetic disease, Fanconi anemia (FA), now attracts broader attention from cancer biologists and basic researchers in the DNA repair and ubiquitin biology fields as well as from hematologists. FA is a chromosome instability syndrome characterized by childhood-onset aplastic anemia, cancer or leukemia susceptibility, and cellular hypersensitivity to DNAcrosslinking agents. Identification of 11 genes for FA has led to progress in the molecular understanding of this disease. FA proteins, including a ubiquitin ligase (FANCL), a monoubiquitinated protein (FANCD2), a helicase (FANCJ/BACH1/BRIP1), and a breast/ovarian cancer susceptibility protein (FANCD1/BRCA2), appear to cooperate in a pathway leading to the recognition and repair of damaged DNA. Molecular interactions among FA proteins and responsible proteins for other chromosome instability syndromes (BLM, NBS1, MRE11, ATM, and ATR) have also been found. Furthermore, inactivation of FA genes has been observed in a wide variety of human cancers in the general population. These findings have broad implications for predicting the sensitivity and resistance of tumors to widely used anticancer DNA crosslinking agents (cisplatin, mitomycin C, and melphalan). Here, we summarize recent progress in the molecular biology of FA and discuss roles of the FA proteins in DNA repair and cancer biology.

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Biallelic Mutations in BRCA1 Cause a New Fanconi Anemia Subtype

Cancer Discovery ◽

10.1158/2159-8290.cd-14-1156 ◽

2014 ◽

Vol 5 (2) ◽

pp. 135-142 ◽

Cited By ~ 185

Author(s):

Sarah L. Sawyer ◽

Lei Tian ◽

Marketta Kähkönen ◽

Jeremy Schwartzentruber ◽

Martin Kircher ◽

...

Keyword(s):

Fanconi Anemia ◽

Biallelic Mutations

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Individuals with FANCM biallelic mutations do not develop Fanconi anemia, but show risk for breast cancer, chemotherapy toxicity and may display chromosome fragility

Genetics in Medicine ◽

10.1038/gim.2017.123 ◽

2017 ◽

Vol 20 (4) ◽

pp. 452-457 ◽

Cited By ~ 32

Author(s):

Irene Catucci ◽

Ana Osorio ◽

Brita Arver ◽

Guido Neidhardt ◽

Massimo Bogliolo ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Chemotherapy ◽

Fanconi Anemia ◽

Chemotherapy Toxicity ◽

Breast Cancer Chemotherapy ◽

Chromosome Fragility ◽

Biallelic Mutations

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Deficiency of UBE2T, the E2 Ubiquitin Ligase Necessary for FANCD2 and FANCI Ubiquitination, Causes FA-T Subtype of Fanconi Anemia

Cell Reports ◽

10.1016/j.celrep.2015.06.014 ◽

2015 ◽

Vol 12 (1) ◽

pp. 35-41 ◽

Cited By ~ 75

Author(s):

Kimberly A. Rickman ◽

Francis P. Lach ◽

Avinash Abhyankar ◽

Frank X. Donovan ◽

Erica M. Sanborn ◽

...

Keyword(s):

Ubiquitin Ligase ◽

Fanconi Anemia

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RAD18-mediated ubiquitination of PCNA activates the Fanconi anemia DNA repair network

The Journal of Cell Biology ◽

10.1083/jcb.201005101 ◽

2010 ◽

Vol 191 (2) ◽

pp. 249-257 ◽

Cited By ~ 67

Author(s):

Liyi Geng ◽

Catherine J. Huntoon ◽

Larry M. Karnitz

Keyword(s):

Ubiquitin Ligase ◽

Fanconi Anemia ◽

Proliferating Cell Nuclear Antigen ◽

E3 Ubiquitin Ligase ◽

Complementation Group ◽

Nuclear Antigen ◽

Group I ◽

Pathway Activation ◽

Cross Links ◽

Proliferating Cell

The Fanconi anemia (FA) network is important for the repair of interstrand DNA cross-links. A key event in FA pathway activation is the monoubiquitylation of the FA complementation group I (FANCI)–FANCD2 (ID) complex by FA complementation group L (FANCL), an E3 ubiquitin ligase. In this study, we show that RAD18, another DNA damage–activated E3 ubiquitin ligase, also participates in ID complex activation by ubiquitylating proliferating cell nuclear antigen (PCNA) on Lys164, an event required for the recruitment of FANCL to chromatin. We also found that monoubiquitylated PCNA stimulates FANCL-catalyzed FANCD2 and FANCI monoubiquitylation. Collectively, these experiments identify RAD18-mediated PCNA monoubiquitination as a central hub for the mobilization of the FA pathway by promoting FANCL-mediated FANCD2 monoubiquitylation.

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