UBE2T, the Fanconi Anemia Core Complex, and FANCD2 Are Recruited Independently to Chromatin: a Basis for the Regulation of FANCD2 Monoubiquitination

Alpi, Arno F.; Langevin, Frédéric; Mosedale, Georgina; Machida, Yuichi; Dutta, Anindya; Patel, Ketan

doi:10.1128/mcb.00504-07

Cited by 82 publications

(96 citation statements)

References 39 publications

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“…Since benign lesions within carcinomas still retain their nuclear staining for FANCD2, this change is specific to the malignant state, and is not a field effect caused by extracellular processes. Since nuclear FANCD2, and specifically monoubiquitylated FANCD2 located in chromatin, 14,15 is generally considered to be its active form in DNA repair processes, the predominant cytoplasmic location of the smaller 155 kDa, non-monoubiquitylated FANCD2 in malignant cells (Figure 2A) suggests that FANCD2-mediated DNA repair would be severely compromised. Interestingly, a recent study also reported the cytoplasmic retention of BRCA1 and RAD51 in a high frequency of sporadic breast cancers, 47 proteins known to associate in the nucleus with FANCD2.…”

Section: Discussionmentioning

confidence: 99%

“…monoubiquitylation, co-localizing with BRCA1, BRCA2 and the recombinase RAD51 in DNA damage inducible foci. [11][12][13][14][15] FANCD2 is primarily regarded as being active within the nucleus and specifically in chromatin, although a cytoplasmic function has recently been proposed. 16 The FA proteins BRCA2/FANCD1 and FANCN/PALB2 are believed to function downstream of, or in parallel with, FANCD2 monoubiquitylation and FANCN and FANCJ/ BRIP1, like BRCA2, are now considered to be breast cancer susceptibility genes.…”

Section: Supported By Grants From the Cancer And Polio Research Fund mentioning

confidence: 99%

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Significance of the Fanconi Anemia FANCD2 Protein in Sporadic and Metastatic Human Breast Cancer

Rudland

Platt-Higgins

Davies

et al. 2010

The American Journal of Pathology

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FANCD2, a pivotal protein in the Fanconi anemia and BRCA pathway/network, is monoubiquitylated in the nucleus in response to DNA damage. This study examines the subcellular location and relationship with prognostic factors and patient survival of FANCD2 in breast cancer. Antibodies to FANCD2 were used to immunocytochemically stain 16 benign and 20 malignant breast specimens as well as 314 primary breast carcinomas to assess its association with subcellular compartment and prognostic factors using Fisher's Exact test or with patient survival over 20 years using Wilcoxon-Gehan statistics. Immunoreactive FANCD2 was found in the nucleus and cytoplasm of all 16 benign tissues, but nuclear staining was lost from a significant 19/20 malignant carcinomas (P < 0.0001). Antibodies to FANCD2 stained the cytoplasm of 196 primary carcinomas, leaving 118 as negatively stained. Negative cytoplasmic staining was significantly associated with positive staining for the metastasis-inducing proteins S100A4, S100P, osteopontin, and AGR2 (P < 0.002). Survival of patients with FANCD2-negative carcinomas was significantly worse (P < 0.0001) than those with positively stained carcinomas, and only 4% were alive at the census date. Multivariate regression analysis identified negative staining for cytoplasmic FANCD2 as the most significant indicator of patient death (P ‫؍‬ 0.001). Thus FANCD2's cytoplasmic loss in the primary carcinomas may allow the selection of cells overexpressing proteins that can induce metastases before surgery. Previous reports have shown that the expression of four proteins that can induce metastasis in experimental rats are highly significantly correlated with each other and separately with early patient death in human breast cancer. These four proteins are S100A4, osteopontin (OPN), AGR2, and S100P.1-4 If their expression is coordinated, then markers of the underlying mechanism should be even more highly correlated with patient demise. One possible coordination mechanism is the generation of an unstable genome by failure of a DNA repair pathway or some other protective mechanism. The majority of familial breast cancer is associated with individuals heterozygous for the BRCA1 or BRCA2 genes that encode proteins important for the repair of DNA double strand breaks and interstrand crosslinks by homologous recombination. 5,6 Biallelic inactivation of BRCA2 results in one form of the cancer-prone syndrome Fanconi anemia (complementation group FA-D1) and the BRCA2/FANCD1 protein operates with 12 other Fanconi anemia (FA) proteins and BRCA1 in a multifaceted response to DNA damage known as the FA/ BRCA tumor suppressor pathway/network. 7-10 Eight of the 13 proteins, together with two FA-associated-proteins, participate in a nuclear core-complex that is required for the monoubiquitylation of FANCD2 and FANCI.

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Section: Discussionmentioning

confidence: 99%

Section: Supported By Grants From the Cancer And Polio Research Fund mentioning

confidence: 99%

Significance of the Fanconi Anemia FANCD2 Protein in Sporadic and Metastatic Human Breast Cancer

Rudland

Platt-Higgins

Davies

et al. 2010

The American Journal of Pathology

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“…The FANCcore complex is an E3 multiprotein complex that contains the E3 RING ligase FANCL, which, in collaboration with the E2 UBE2T, catalyzes the transfer of a monoubiquitin to the downstream targets FANCD2 and FANCI (Alpi et al, 2007;Alpi et al, 2008;Machida et al, 2006). FANCcore complex partners, including FANCA, FANCB, FANCC, FANCE, FANCF, FANCG, FANCL and FANCM, and its two identified targets constitute the upper portion of a signaling cascade referred to as the FANC pathway.…”

Section: Introductionmentioning

confidence: 99%

Proteomic analysis unveils a FANCA-modulated neddylation pathway involved in CXCR5 membrane targeting and cell mobility

Renaudin

Guervilly

Aoufouchi

et al. 2014

Journal of Cell Science

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The aim of this study was to identify novel substrates of the FANCcore complex, the inactivation of which leads to the genetic disorder Fanconi anemia, which is associated with bone marrow failure, developmental abnormalities and a predisposition to cancer. Eight FANC proteins participate in the nuclear FANCcore complex, which functions as an E3 ubiquitin-ligase that monoubiquitylates FANCD2 and FANCI in response to replicative stress. Here, we use mass spectrometry to compare proteins from FANCcore-complexdeficient cells to those of rescued control cells after treatment with hydroxyurea, an inducer of FANCD2 monoubiquitylation. FANCD2 and FANCI appear to be the only targets of the FANCcore complex. We identify other proteins that are post-translationally modified in a FANCA-or FANCC-dependent manner. The majority of these potential targets localize to the cell membrane. Finally, we demonstrate that (a) the chemokine receptor CXCR5 is neddylated; (b) FANCA but not FANCC appears to modulate CXCR5 neddylation through an unknown mechanism; (c) CXCR5 neddylation is involved in targeting the receptor to the cell membrane; and (d) CXCR5 neddylation stimulates cell migration and motility. Our work has uncovered a pathway involving FANCA in neddylation and cell motility.

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“…[29][30][31] The remaining three FA proteins are breast cancer susceptibility related genes (BRCA) FANCD1/ BRCA2, FANCJ, FANCN, which are thought to function downstream of FANCD2/I. 28,[32][33][34][35] Although the FA pathway is widely hypothesized to participate in DNA repair and maintenance of genomic stability, its precise biochemical role(s) are unknown. The FA pathway is implicated in the repair of DNA interstrand crosslinks and fork-stalling lesions.…”

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confidence: 99%

Rad18 E3 ubiquitin ligase activity mediates Fanconi anemia pathway activation and cell survival following DNA Topoisomerase 1 inhibition

Palle¹,

Vaziri²

2011

Cell Cycle

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Camptothecin (Cpt) and related chemotherapeutic drugs induce formation of DNA topoisomerase I (top1) covalent or cleavage complexes (top1ccs) that block leading-strand DNA synthesis and elicit DNA Double Stranded Breaks (DSB) during S phase. the Fanconi Anemia (FA) pathway is implicated in tolerance of Cpt-induced DNA damage yet the mechanism of FA pathway activation by top1 poisons has not been studied. We show here that the FA core complex protein FANCA and monoubiquitinated FANCD2 (an effector of the FA pathway) are rapidly mobilized to chromatin in response to Cpt treatment in several human cancer cell lines and untransformed primary human dermal fibroblasts. FANCD2 depletion using siRNA leads to impaired recovery from Cpt-induced inhibition or DNA synthesis, persistence of γH2AX (a DSB marker) and reduced cell survival following Cpt treatment. the e3 ubiquitin ligase Rad18 is necessary for Cpt-induced recruitment of FANCA and FANCD2 to chromatin. Moreover, Rad18-depletion recapitulates the DNA synthesis and survival defects of FANCD2-deficiency in Cpt-treated cells. It is well-established that Rad18 promotes FA pathway activation and DNA damage tolerance in response to bulky DNA lesions via a mechanism involving pCNA monoubiquitination. In contrast, pCNA monoubiquitination is not involved in Rad18-mediated FA pathway activation or cell survival following acquisition of Cpt-induced DSB. Moreover, while Rad18 is implicated in recombinational repair of DSB via an e3 ligase-independent mechanism, we demonstrate that Rad18 e3 ligase activity is essential for appropriate FA pathway activation and DNA damage tolerance after Cpt treatment. taken together, our results define a novel pathway of Rad18-dependent DSB repair that is dissociable from known Rad18-mediated DNA repair mechanisms based on its independence from pCNA ubiquitination and requirement for e3 ligase activity. MMS, methyl methanesulfonate; NBS1, nijmegen breakage syndrome protein1; NHEJ, non-homologous end joining; PBS, phosphate-buffered saline; PCNA, proliferating cell nuclear antigen; Polη, DNA polymerases eta; Polι, DNA polymerases iota; Polκ, DNA polymerases kappa; RFC, replication factor C; RPA, replication protein A; RT, room temperature; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; siRNA, small interference RNA; ssDNA, single-stranded DNA; TBE, tris-boric acid-EDTA (buffer); TLS, translesion DNA synthesis; Top1, DNA topoisomerase I; Top1cc, DNA-topoisomerase I covalent or cleavage complex; Usp1, ubiquitin-specific processing protease 1; UV, ultra violet radiation; WT, wild type ©2 0 1 1 L a n d e s B i o s c i e n c e . D o n o t d i s t r i b u t e .1626 Cell Cycle Volume 10 Issue 10 with TLS, ensuring appropriate processing of bulky DNA lesions. FA is a cancer predisposition syndrome in humans. To date, at least 13 complementation groups of FA have been identified (designated 'FANC' A-M). The FA proteins (FANCs) are thought to work cooperatively in a common pathway to repair interstrand crosslinks. 28 The eight-prote...

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UBE2T, the Fanconi Anemia Core Complex, and FANCD2 Are Recruited Independently to Chromatin: a Basis for the Regulation of FANCD2 Monoubiquitination

Cited by 82 publications

References 39 publications

Significance of the Fanconi Anemia FANCD2 Protein in Sporadic and Metastatic Human Breast Cancer

Significance of the Fanconi Anemia FANCD2 Protein in Sporadic and Metastatic Human Breast Cancer

Proteomic analysis unveils a FANCA-modulated neddylation pathway involved in CXCR5 membrane targeting and cell mobility

Rad18 E3 ubiquitin ligase activity mediates Fanconi anemia pathway activation and cell survival following DNA Topoisomerase 1 inhibition

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