Biallelic mutations in the ubiquitin ligase RFWD3 cause Fanconi anemia

Abstract

The WD40-containing E3 ubiquitin ligase RFWD3 has been recently linked to the repair of DNA damage by homologous recombination (HR). Here we have shown that an RFWD3 mutation within the WD40 domain is connected to the genetic disease Fanconi anemia (FA). An individual presented with congenital abnormalities characteristic of FA. Cells from the patient carrying the compound heterozygous mutations c.205_206dupCC and c.1916T>A in RFWD3 showed increased sensitivity to DNA interstrand cross-linking agents in terms of increased chromosomal breakage, reduced survival, and cell cycle arrest in G 2 phase. The cellular phenotype was mirrored in genetically engineered human and avian cells by inactivation of RFWD3 or introduction of the patient-derived missense mutation, and the phenotype was rescued by expression of wild-type RFWD3 protein. HR was disrupted in RFWD3-mutant cells as a result of impaired relocation of mutant RFWD3 to chromatin and defective physical interaction with replication protein A. Rfwd3 knockout mice appear to have increased embryonic lethality, are subfertile, show ovarian and testicular atrophy, and have a reduced lifespan resembling that of other FA mouse models. Although RFWD3 mutations have thus far been detected in a single child with FA, we propose RFWD3 as an FA gene, FANCW, supported by cellular paradigm systems and an animal model.

Fanconi Anemia Signaling and Cancer

“…Importantly, FANCM-FAAP24 was also found to interact with ATR-ATRIP and HCLK2 in FA -A, C, and G cells [69], this observation is distinct from the role of FANCM in forming the FA complex E3, which activates FANCD2/I [12]. Aside from FANCM affecting ATR activation, the newly identified FANCW also plays an important role in ATR functions, at which point can ubiquitinate RPA and modulate the functions of RPA [4, 5]. In addition, the relationship between the FA proteins and ATR may be mutual and of equally importance.…”

p53-TP53-Induced Glycolysis Regulator Mediated Glycolytic Suppression Attenuates DNA Damage and Genomic Instability in Fanconi Anemia Hematopoietic Stem Cells

“…Fanconi anemia (FA) is a rare inherited disease characterized by bone marrow (BM) failure and high risk of neoplasia including leukemia from dysfunctional hematopoietic stem cells (HSCs; [1,2]). FA is genetically heterogeneous with at least 22 genes (FANCA-W) identified thus far [1][2][3][4][5][6][7][8][9][10][11]. The hematologic complications of FA are nearly universal among patients with FA and defects in HSCs have been postulated as the root cause of BM failure and leukemia in FA [6,12].…”

Target highlights in CASP13: Experimental target structures through the eyes of their authors

“…2,3 Evidence has shown that the WDR domain is primarily responsible for the functional interactions that allow RFWD3 to maintain genomic stability. 4,5 Furthermore, heritable mutations, particularly an Ile639Lys point mutation within the WDR domain, may lead to the rare genomic instability disease known as Fanconi anemia (FA), 6 thereby implicating RFWD3 as a potential FA-associated gene (alias: FANCW). [4][5][6] Until recently, the biochemical characterization of RFWD3 has lacked complementary highresolution structural information.…”

“…4,5 Furthermore, heritable mutations, particularly an Ile639Lys point mutation within the WDR domain, may lead to the rare genomic instability disease known as Fanconi anemia (FA), 6 thereby implicating RFWD3 as a potential FA-associated gene (alias: FANCW). [4][5][6] Until recently, the biochemical characterization of RFWD3 has lacked complementary highresolution structural information. Here, we discuss the 1.8 Å resolution X-ray crystal structure of the C-terminal WDR domain of human RFWD3 ( Table 1, Target: T0954; PDB: 6CVZ).…”

“…ates the last eight helices(3)(4)(5)(6)(7)(8)(9)(10). Connecting the helices are the HVSs that form short β-turn loops at both ends of the domain.…”