scholarly journals Fanconi anemia proteins counteract the implementation of the oncogene-induced senescence program

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Anne Helbling-Leclerc ◽  
Françoise Dessarps-Freichey ◽  
Caroline Evrard ◽  
Filippo Rosselli
Keyword(s):  
Fanconi Anemia ◽  
Bone Marrow Failure ◽  
Cathepsin L ◽  
Loss Of Function ◽  
Developmental Abnormalities ◽  
Cellular Models ◽  
Senescent Phenotype ◽  
Pathway Activation ◽  
Oncogene Activation ◽  
Senescence Program

AbstractFanconi Anemia (FA), due to the loss-of-function of the proteins that constitute the FANC pathway involved in DNA replication and genetic stability maintainance, is a rare genetic disease featuring bone marrow failure, developmental abnormalities and cancer predisposition. Similar clinical stigmas have also been associated with alterations in the senescence program, which is activated in physiological or stress situations, including the unscheduled, chronic, activation of an oncogene (oncogene induced senescence, OIS). Here, we wanted to determine the crosstalk, if any, between the FANC pathway and the OIS process. OIS was analyzed in two known cellular models, IMR90-hTERT/ER:RASG12V and WI38-hTERT/ER:GFP:RAF1, harboring 4-hydroxytamoxifen-inducible oncogenes. We observed that oncogene activation induces a transitory increase of both FANCA and FANCD2 as well as FANCD2 monoubiquitination, readout of FANC pathway activation, followed by their degradation. FANCD2 depletion, which leads to a pre-senescent phenotype, anticipates OIS progression. Coherently, FANCD2 overexpression or inhibition of its proteosomal-dependent degradation slightly delays OIS progression. The pro-senescence protease cathepsin L, which activation is anticipated during OIS in FANCD2-depleted cells, also participates to FANCD2 degradation. Our results demonstrate that oncogene activation is first associated with FANCD2 induction and activation, which may support initial cell proliferation, followed by its degradation/downregulation when OIS proceeds.

scholarly journals Fanconi anemia A protein participates in nucleolar homeostasis maintenance and ribosome biogenesis

2021 ◽  
Vol 7 (1) ◽  
pp. eabb5414
Author(s):  
Anna Gueiderikh ◽  
Frédérique Maczkowiak-Chartois ◽  
Guillaume Rouvet ◽  
Sylvie Souquère-Besse ◽  
Sébastien Apcher ◽  
...  
Keyword(s):  
Dna Damage ◽  
Fanconi Anemia ◽  
Stress Responses ◽  
Ribosome Biogenesis ◽  
Bone Marrow Failure ◽  
Loss Of Function ◽  
Independent Manner ◽  
Ribosomal Subunits ◽  
Nucleolar Proteins ◽  
Nucleolar Stress

Fanconi anemia (FA), the most common inherited bone marrow failure and leukemia predisposition syndrome, is generally attributed to alterations in DNA damage responses due to the loss of function of the DNA repair and replication rescue activities of the FANC pathway. Here, we report that FANCA deficiency, whose inactivation has been identified in two-thirds of FA patients, is associated with nucleolar homeostasis loss, mislocalization of key nucleolar proteins, including nucleolin (NCL) and nucleophosmin 1 (NPM1), as well as alterations in ribosome biogenesis and protein synthesis. FANCA coimmunoprecipitates with NCL and NPM1 in a FANCcore complex–independent manner and, unique among the FANCcore complex proteins, associates with ribosomal subunits, influencing the stoichiometry of the translational machineries. In conclusion, we have identified unexpected nucleolar and translational consequences specifically associated with FANCA deficiency that appears to be involved in both DNA damage and nucleolar stress responses, challenging current hypothesis on FA physiopathology.

scholarly journals HES1 is a novel interactor of the Fanconi anemia core complex

Blood ◽  
2008 ◽  
Vol 112 (5) ◽  
pp. 2062-2070 ◽  
Author(s):  
Cédric S. Tremblay ◽  
Feng F. Huang ◽  
Ouassila Habi ◽  
Caroline C. Huard ◽  
Chantal Godin ◽  
...  
Keyword(s):  
Fanconi Anemia ◽  
Bone Marrow Failure ◽  
Developmental Pathway ◽  
Core Complex ◽  
Interacting Protein ◽  
Developmental Abnormalities ◽  
Protein Protein Interaction ◽  
The Core ◽  
Cross Links ◽  
Enhancer Of Split

Abstract Fanconi anemia (FA) proteins are thought to play a role in chromosome stability and repair of DNA cross-links; however, these functions may not fully explain the developmental abnormalities and bone marrow failure that are characteristic of FA individuals. Here we associate the FA proteins with the Notch1 developmental pathway through a direct protein-protein interaction between the FA core complex and the hairy enhancer of split 1 (HES1). HES1 interaction with FA core complex members is dependent on a functional FA pathway. Cells depleted of HES1 exhibit an FA-like phenotype that includes cellular hypersensitivity to mitomycin C (MMC) and lack of FANCD2 monoubiquitination and foci formation. HES1 is also required for proper nuclear localization or stability of some members of the core complex. Our results suggest that HES1 is a novel interacting protein of the FA core complex.

scholarly journals Enhanced frequency of sister chromatid exchanges induced by diepoxybutane is specific characteristic of Fanconi anemia cellular phenotype

Genetika ◽  
2013 ◽  
Vol 45 (2) ◽  
pp. 393-403 ◽  
Author(s):  
Ivana Joksic ◽  
Sandra Petrovic ◽  
Andreja Leskovac ◽  
Jelena Filipovic ◽  
Marija Guc-Scekic ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Fanconi Anemia ◽  
Sister Chromatid ◽  
Cancer Susceptibility ◽  
Bone Marrow Failure ◽  
Sister Chromatid Exchanges ◽  
Healthy Controls ◽  
Developmental Abnormalities ◽  
Radiation Induced ◽  
Chromatid Exchanges

Fanconi anemia (FA) is a rare genetically heterogeneous disease characterized by developmental abnormalities, progressive bone marrow failure, and cancer susceptibility. We examined spontaneous, diepoxybutane (DEB)- induced and radiation-induced sister chromatid exchanges (SCEs) in wholeblood lymphocyte cultures of bone marrow failure (BMF) patients including Fanconi anemia, mothers of affected individuals, and healthy controls. The baseline frequency of SCE in FA cells was similar to that observed in controls. However, in response to DEB SCE frequencies in FA patients and their mothers were significantly increased compared to both non-FA BMF families and healthy controls. In response to ionizing radiation, cells displayed increased frequency of SCE, but no differences between FA patients and non-FA BMF patients were seen. Our data confirm and expand previous findings by showing that SCE induced by DEB can be used as an adjunct diagnostic test not only for FA patients, but also for female heterozygous carriers, at least for complementation groups FANCA and FANCD2.

Clinical Characteristics of Patients with Fanconi Anemia in Complementation Group J.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3769-3769 ◽  
Author(s):  
Angeles Dasi-Carpio ◽  
Jose A. Casado ◽  
Guillermo Sanz ◽  
Paula Rio ◽  
Maria-Teresa Santos ◽  
...  
Keyword(s):  
Short Stature ◽  
Fanconi Anemia ◽  
Clinical Course ◽  
Lentiviral Vector ◽  
Cancer Susceptibility ◽  
Bone Marrow Failure ◽  
Skin Pigmentation ◽  
Complementation Group ◽  
Developmental Abnormalities ◽  
Female Male Female

Abstract Fanconi anemia (FA) is an inherited disorder characterized by chromosomal instability, developmental abnormalities, progressive bone-marrow failure, and cancer susceptibility. To date, twelve complementation groups have been described for FA (A, B, C, D1, D2, E, F, G, I, J, L and M) and eleven associated genes have already been identified (FANCA, FANCB, FANCC, FANCD1/BRCA2, FANCD2, FANCE, FANCF, FANCG/XRCC9, BRIP1/FANCJ, FANCL and FANCM/Hef). The FA complementation group J (FA-J) was first reported in 2004. FA-A is the most common group, accounting for approximately 65% of all affected individuals while FA-J is a rare group, comprising only 1.6% of all FA patients in the International Fanconi Anemia Registry (IFAR). To the best of our knowledge 11 FA-J patients have been reported in the IFAR. Patients with Fanconi anemia show extreme clinical heterogeneity. Thus, it is of interest to associate the phenotype with the complemantation group, since this could provide predictive information on clinical outcome. So far, in the Hospital La Fe, 4 patients (two siblings and two unrelated patients) belong to scarce FA-J complementation group. This represents the 4.5 % of the FA patients included in the Spanish Fanconi Anemia Registry. Subtyping was done by viral complementation. Cells from these patients were not complemented with a battery of vectors with FA genes (FANCA, FANCC, FANCG, FANCE, FANCF, and FANCD2). However, the characteristic cellular hypersensitivity to mitomycin C was reverted when a lentiviral vector encoding for BRIP1/FANCJ was used. Moreover, mutational studies confirmed the subtyping of FA-J in these Fanconi patients. The clinical course of the 4 FA-J patienst are shown in the Table. In conclusion, its noteworthy the high frequency of the FA-J genotype in Spain. The 75% of our patients have major congenital malformations and severe endocrinopathy. All our patients had short stature and 50% were mycrosomic. None of our patients had skin abnormalities, although this is present in 80% of the patients in other registries. Although two of these patients developed aplasia very early (5 years old), FA-J siblings developed severe aplasia at the age of 20 years but none of them developed leukemia or solid tumors. Clinical course of patients Patient 1* 2* 3 4 * Sibling sex male female male female Aplasia onset (years) 24 20.5 4.3 3.5 Survival (years) 28 31.5 6 alive Short stature Yes Yes Yes Yes Microsomy No Yes Yes No Skin pigmentation No No No No Eyes abnormalities No No microphthalmia Epiphora Skeletal malformations Syndactyly No Abnormal thumbs Abnormal thumbs Kidneys Agenesis, left No No Horseshoe Endocrinopathy failure puberty not available hypotiroidism hypopituitarysm Gastrointestinal malformations No No Esophageal atresia Onphalocele Malignacies No No No No Family cancers Not available Not available Liver Lung, brains and breast

scholarly journals A Dutch Fanconi AnemiaFANCCFounder Mutation in Canadian Manitoba Mennonites

Anemia ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Yne de Vries ◽  
Nikki Lwiwski ◽  
Marieke Levitus ◽  
Bertus Kuyt ◽  
Sara J. Israels ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Fanconi Anemia ◽  
Bone Marrow Failure ◽  
Complementation Group ◽  
Cross Linking ◽  
Founder Mutations ◽  
Developmental Abnormalities ◽  
Dna Instability ◽  
Common Founder ◽  
Complementation Groups

Fanconi anemia (FA) is a recessive DNA instability disorder associated with developmental abnormalities, bone marrow failure, and a predisposition to cancer. Based on their sensitivity to DNA cross-linking agents, FA cells have been assigned to 15 complementation groups, and the associated genes have been identified. Founder mutations have been found in different FA genes in several populations. The majority of Dutch FA patients belongs to complementation group FA-C. Here, we report 15 patients of Dutch ancestry and a large Canadian Manitoba Mennonite kindred carrying theFANCCc.67delG mutation. Genealogical investigation into the ancestors of the Dutch patients shows that these ancestors lived in four distinct areas in The Netherlands. We also show that the Dutch and Manitoba MennoniteFANCCc.67delG patients share the same haplotype surrounding this mutation, indicating a common founder.

scholarly journals Cell cycle–dependent chromatin loading of the Fanconi anemia core complex by FANCM/FAAP24

Blood ◽  
2008 ◽  
Vol 111 (10) ◽  
pp. 5215-5222 ◽  
Author(s):  
Jung Min Kim ◽  
Younghoon Kee ◽  
Allan Gurtan ◽  
Alan D. D'Andrea
Keyword(s):  
Cell Cycle ◽  
Fanconi Anemia ◽  
Cancer Susceptibility ◽  
Bone Marrow Failure ◽  
Core Complex ◽  
Developmental Abnormalities ◽  
Repair Protein ◽  
Dna Repair Protein ◽  
Human Ortholog ◽  
Cycle Dependent

Abstract Fanconi anemia (FA) is a genetic disease characterized by congenital abnormalities, bone marrow failure, and cancer susceptibility. A total of 13 FA proteins are involved in regulating genome surveillance and chromosomal stability. The FA core complex, consisting of 8 FA proteins (A/B/C/E/F/G/L/M), is essential for the monoubiquitination of FANCD2 and FANCI. FANCM is a human ortholog of the archaeal DNA repair protein Hef, and it contains a DEAH helicase and a nuclease domain. Here, we examined the effect of FANCM expression on the integrity and localization of the FA core complex. FANCM was exclusively localized to chromatin fractions and underwent cell cycle–dependent phosphorylation and dephosphorylation. FANCM-depleted HeLa cells had an intact FA core complex but were defective in chromatin localization of the complex. Moreover, depletion of the FANCM binding partner, FAAP24, disrupted the chromatin association of FANCM and destabilized FANCM, leading to defective recruitment of the FA core complex to chromatin. Our results suggest that FANCM is an anchor required for recruitment of the FA core complex to chromatin, and that the FANCM/FAAP24 interaction is essential for this chromatin-loading activity. Dysregulated loading of the FA core complex accounts, at least in part, for the characteristic cellular and developmental abnormalities in FA.

scholarly journals Fanconi anemia proteins are required to maintain nucleolar homeostasis

2019 ◽  
Author(s):  
Anna Gueiderikh ◽  
Guillaume Rouvet ◽  
Sylvie Souquère-Besse ◽  
Sébastien Apcher ◽  
Jean-Jacques Diaz ◽  
...  
Keyword(s):  
Dna Damage ◽  
Fanconi Anemia ◽  
Ribosome Biogenesis ◽  
Bone Marrow Failure ◽  
Rrna Processing ◽  
Loss Of Function ◽  
Rdna Transcription ◽  
Inhibitory Protein ◽  
Growth Inhibitory ◽  
Dna Damage Responses

AbstractThe majority of inherited bone marrow failure (iBMF) syndromes are associated to nucleolar and/or ribosomal abnormalities, but Fanconi anemia (FA), the most common iBMF, is attributed to alterations in DNA damage responses. However, the involvement, if any, of the FA (FANC) proteins in the maintenance of nucleolar functions and/or ribosome biogenesis is yet unexplored. Here, we report that FANC pathway loss-of-function is associated to a loss of the nucleolar homeostasis, demonstrating increased rDNA rearrangements, accumulation of nucleolar DNA damage, nucleolar protein mislocalization, and a p53-independent induction of the growth inhibitory protein p21. Moreover, specifically associated to FANCA loss-of-function, which is responsible for approximately 65% of FA cases, we observed reduced rDNA transcription and rRNA processing as well as alteration in protein synthesis and polysome profiles. Thus, we have identified nucleolar consequences associated with FANC pathway deficiency, challenging current hypothesis on the physiopathology of FA.

scholarly journals Loss of the homologous recombination generad51leads to Fanconi anemia-like symptoms in zebrafish

2017 ◽  
Vol 114 (22) ◽  
pp. E4452-E4461 ◽  
Author(s):  
Jan Gregor Botthof ◽  
Ewa Bielczyk-Maczyńska ◽  
Lauren Ferreira ◽  
Ana Cvejic
Keyword(s):  
Bone Marrow ◽  
Homologous Recombination ◽  
Fanconi Anemia ◽  
Bone Marrow Failure ◽  
Marrow Cell ◽  
Tumor Development ◽  
Dominant Negative ◽  
Loss Of Function ◽  
Hematopoietic Stem ◽  
Inflammatory Stress

RAD51 is an indispensable homologous recombination protein, necessary for strand invasion and crossing over. It has recently been designated as a Fanconi anemia (FA) gene, following the discovery of two patients carrying dominant-negative mutations. FA is a hereditary DNA-repair disorder characterized by various congenital abnormalities, progressive bone marrow failure, and cancer predisposition. In this report, we describe a viable vertebrate model ofRAD51loss. Zebrafishrad51loss-of-function mutants developed key features of FA, including hypocellular kidney marrow, sensitivity to cross-linking agents, and decreased size. We show that some of these symptoms stem from both decreased proliferation and increased apoptosis of embryonic hematopoietic stem and progenitor cells. Comutation ofp53was able to rescue the hematopoietic defects seen in the single mutants, but led to tumor development. We further demonstrate that prolonged inflammatory stress can exacerbate the hematological impairment, leading to an additional decrease in kidney marrow cell numbers. These findings strengthen the assignment ofRAD51as a Fanconi gene and provide more evidence for the notion that aberrant p53 signaling during embryogenesis leads to the hematological defects seen later in life in FA. Further research on this zebrafish FA model will lead to a deeper understanding of the molecular basis of bone marrow failure in FA and the cellular role of RAD51.

Aberrant activation of stress-response pathways leads to TNF-α oversecretion in Fanconi anemia

Blood ◽  
2008 ◽  
Vol 111 (4) ◽  
pp. 1913-1923 ◽  
Author(s):  
Delphine Briot ◽  
Gaëtane Macé-Aimé ◽  
Frédéric Subra ◽  
Filippo Rosselli
Keyword(s):  
Bone Marrow ◽  
Fanconi Anemia ◽  
Cytoplasmic Membrane ◽  
Bone Marrow Failure ◽  
Mapk Signaling ◽  
High Rate ◽  
Loss Of Function ◽  
Aberrant Expression ◽  
Tnf Α ◽  
The Matrix

Fanconi anemia (FA), an inherited syndrome that associates bone marrow failure, cancer predisposition, and genetic instability, is characterized by an overproduction of the myelosuppressive cytokine TNF-α through unknown mechanisms. We demonstrate here that FANC pathway loss-of-function results in the aberrant activation of 2 major stress-signaling pathways: NF-κB and MAPKs. These responses are independent on TNF-α expression. On the contrary, inhibition of the MAPK pathways normalizes TNF-α oversecretion in FA. Moreover, our data show that the overexpression of the matrix metalloproteinase MMP-7 is the key event directly responsible for the high rate of TNF-α shedding and release from the cytoplasmic membrane in FA. TNF-α overproduction is, indeed, normalized by MMP-7 inhibition. Finally, MAPK inhibition impacts on MMP-7 overexpression. Evidence is provided of the existence of a linear pathway in which FANC mutations activate MAPK signaling that induces MMP-7 overexpression leading, in fine, to TNF-α oversecretion. TNF-α may, in turn, sustain or amplify both MAPKs and NF-κB activation. Aberrant expression or activity of NF-κB and/or MAPKs has been already involved in bone marrow failure and leukemia, and their inhibition offered clinical benefit for patients. In conclusion, our data provide a strong rationale for new clinical trials on FA patients.

Disparate Requirements for the Phosphorylation of Distinct Sites on the Fanconi Anemia Group I Protein

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 358-358
Author(s):  
Ronald S. Cheung ◽  
Maria Castella ◽  
Toshiyasu Taniguchi
Keyword(s):  
Fanconi Anemia ◽  
Bone Marrow Failure ◽  
Human Cells ◽  
The Other ◽  
Phosphorylation Sites ◽  
Core Complex ◽  
Blood Disorder ◽  
Group I ◽  
Pathway Activation ◽  
Anemia Group

Abstract Fanconi Anemia (FA) is a blood disorder characterized by bone marrow failure, predisposition to hematologic malignancy and sensitivity to interstrand crosslinking agents. Patients with FA carry inherited mutations in any one of at least 16 known Fanconi Anemia Group (FANC) proteins that coordinate to function in a DNA repair pathway (the FA pathway). The activation of this pathway centers on two of these, Fanconi Anemia Group D2 protein (FANCD2) and Fanconi Anemia Group I protein (FANCI), which must undergo both phosphorylation and ubiquitination in order for the pathway to function properly. The latter is catalyzed by the FA core complex ubiquitin ligase, which is composed of 8 other FANC proteins. Previous studies suggest that, in response to DNA damage, FANCI is phosphorylated at multiple sites within its evolutionarily conserved SQ cluster domain (SCD). This process is essential for activation of the canonical FA pathway. Failure of FANCI to phosphorylate inhibits FANCD2 ubiquitination, FANCD2 foci formation and cellular resistance to interstrand crosslinkers. However, while FANCI phosphorylation is important for the FA pathway to function, little is known about how this phosphorylation is regulated. Studies on the regulation of FANCI phosphorylation have largely been limited to chicken DT40 cells. Furthermore, the detection of FANCI phosphorylation has been restricted to an electrophoretic mobility-based method, which provides little information on the biology of specific phosphorylation sites. The objective of our work is to better understand the precise regulation of FANCI SCD phosphorylation, in human cells, at sites that have been established to be functionally significant. By performing mass spectrometry on immunoprecipitated human FANCI protein, we established that the human FANCI SCD is indeed phosphorylated on at least two sites. Each of these sites have been found, through mutagenesis studies, to be involved in FA pathway activation. These two sites have also been implicated, through structural studies, in promoting a stable interaction between FANCI and FANCD2. Using this information, we designed immunogenic phospho-peptides to generate antibodies that specifically detect the phosphorylation of each of these two sites. We used these FANCI phospho-antibodies, together with genetically manipulated human cell culture systems, to study factors that modulate FANCI phosphorylation in the context of the human FA pathway. We first established that these antibodies can be used for both immunoblot and immunofluorescence applications. With immunoblot analysis of cells treated with mitomycin C, we made the interesting observation that the phosphorylation of one of the FANCI sites occurred predominantly in the non-ubiquitinated form of the protein, while the other site was phosphorylated predominantly in the ubiquitinated form. This suggested that the phosphorylation of two distinct FANCI sites occurs at different steps of FA pathway activation. By performing siRNA depletion and biochemical experiments in cultured human cells, we found that the phosphorylation of both sites is at least partially dependent on the Ataxia Telangiectasia and Rad 3 related (ATR) kinase. Surprisingly, we found that only one of these sites could be phosphorylated without prior FANCI/D2 ubiquitination. Phosphorylation of the other site was dependent on both FANCI/D2 ubiquitination and the FA core complex. Therefore, contrary to previous models, we found that both ubiquitination-dependent and -independent phosphorylation sites exist within the FANCI SCD. Different FANCI phosphorylation sites that contribute to FA pathway activation therefore have disparate requirements for their phosphorylation. Until now, studies on the regulation of FANCI phosphorylation have been limited by the lack of available phospho-specific FANCI antibodies. By developing antibodies that can specifically detect the phosphorylation of distinct sites within the functionally important SCD of FANCI, we have established new and critical reagents that provide additional insight into how the human FA pathway is activated. Our results suggest a novel model of FA pathway activation that involves a dynamic interplay between FANCI phosphorylation and FANCI/D2 ubiquitination, and reveal that activation of the FA pathway by FANCI phosphorylation is more complex a process than previously thought. Disclosures No relevant conflicts of interest to declare.

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