An ALS-associated mutation in the FUS 3′-UTR disrupts a microRNA–FUS regulatory circuitry

Modigliani, Stefano Dini; Morlando, Mariangela; Errichelli, Lorenzo; Sabatelli, Mario; Bozzoni, Irene

doi:10.1038/ncomms5335

Cited by 104 publications

(94 citation statements)

References 31 publications

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“…Therefore, it is tempting to speculate that mutations disrupting the protein structure may not only be completely responsible for the disease but also changes in FUS protein levels. In fact, mutations in the 3′-UTR of the FUS gene that lead to increased FUS levels through an altered feed-forward regulatory loop, are associated with ALS (Dini Modigliani et al, 2014), suggesting that FUS has to be properly regulated to avoid pathology. Non-neuronal cells, which do not recapitulate the complexity of neurons, as well as neurons have also been extensively used in vitro as valuable tools to analyze cell specific responses but only by overexpressing WT or mFUS (Dormann et al, 2010; Bentmann et al, 2012; Jäckel et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

FUS Mislocalization and Vulnerability to DNA Damage in ALS Patients Derived hiPSCs and Aging Motoneurons

Higelin

Demestre

Putz

et al. 2016

Front. Cell. Neurosci.

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Mutations within the FUS gene (Fused in Sarcoma) are known to cause Amyotrophic Lateral Sclerosis (ALS), a neurodegenerative disease affecting upper and lower motoneurons. The FUS gene codes for a multifunctional RNA/DNA-binding protein that is primarily localized in the nucleus and is involved in cellular processes such as splicing, translation, mRNA transport and DNA damage response. In this study, we analyzed pathophysiological alterations associated with ALS related FUS mutations (mFUS) in human induced pluripotent stem cells (hiPSCs) and hiPSC derived motoneurons. To that end, we compared cells carrying a mild or severe mFUS in physiological- and/or stress conditions as well as after induced DNA damage. Following hyperosmolar stress or irradiation, mFUS hiPS cells recruited significantly more cytoplasmatic FUS into stress granules accompanied by impaired DNA-damage repair. In motoneurons wild-type FUS was localized in the nucleus but also deposited as small punctae within neurites. In motoneurons expressing mFUS the protein was additionally detected in the cytoplasm and a significantly increased number of large, densely packed FUS positive stress granules were seen along neurites. The amount of FUS mislocalization correlated positively with both the onset of the human disease (the earlier the onset the higher the FUS mislocalization) and the maturation status of the motoneurons. Moreover, even in non-stressed post-mitotic mFUS motoneurons clear signs of DNA-damage could be detected. In summary, we found that the susceptibility to cell stress was higher in mFUS hiPSCs and hiPSC derived motoneurons than in controls and the degree of FUS mislocalization correlated well with the clinical severity of the underlying ALS related mFUS. The accumulation of DNA damage and the cellular response to DNA damage stressors was more pronounced in post-mitotic mFUS motoneurons than in dividing hiPSCs suggesting that mFUS motoneurons accumulate foci of DNA damage, which in turn might be directly linked to neurodegeneration.

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Section: Discussionmentioning

confidence: 99%

FUS Mislocalization and Vulnerability to DNA Damage in ALS Patients Derived hiPSCs and Aging Motoneurons

Higelin

Demestre

Putz

et al. 2016

Front. Cell. Neurosci.

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“…Over the last decade, TDP-43, FUS, Matrin-3, VCP and several other RNA-binding proteins have been found to be linked with causing ALS pathogenesis suggesting that disease-causing mutations perturbing RNA metabolism might be central to causing ALS pathogenesis [26, 51, 52, 64, 68]. This is supported by recent studies showing that ALS-causing mutations in RNA binding proteins cause defects in RNA splicing, stability, transcription, mRNA processing, translation and transport, which may lead to gross functional impairments in several key biological pathways [3, 5–7, 10, 18–23, 25, 30, 31, 33, 34, 60, 61, 63, 65, 66]. …”

Section: Introductionmentioning

confidence: 92%

Pur-alpha regulates cytoplasmic stress granule dynamics and ameliorates FUS toxicity

et al. 2016

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Amyotrophic lateral Sclerosis is characterized by progressive loss of motor neurons in the brain and spinal cord. Mutations in several genes, including FUS, TDP43, Matrin 3, hnRNPA2 and other RNA binding proteins, have been linked to ALS pathology. Recently, Pur-alpha a DNA/RNA binding protein was found to bind to C9orf72 repeat expansions and could possibly play a role in the pathogenesis of ALS. When overexpressed, Pur-alpha mitigates toxicities associated with Fragile X tumor ataxia syndrome (FXTAS) and C9orf72 repeat expansion diseases in Drosophila and mammalian cell culture models. However, the function of Pur-alpha in regulating ALS pathogenesis has not been fully understood. We identified Pur-alpha as a novel component of cytoplasmic stress granules (SGs) in ALS patient cells carrying disease-causing mutations in FUS. When cells were challenged with stress, we observed that Pur-alpha co-localized with mutant FUS in ALS patient cells and became trapped in constitutive SGs. We also found that FUS physically interacted with Pur-alpha in mammalian neuronal cells. Interestingly, shRNA mediated knock down of endogenous Pur-alpha significantly reduced formation of cytoplasmic stress granules in mammalian cells suggesting that Pur-alpha is essential for the formation of SGs. Furthermore, ectopic expression of Pur-alpha blocked cytoplasmic mislocalization of mutant FUS and strongly suppressed toxicity associated with mutant FUS expression in primary motor neurons. Our data emphasizes the importance of stress granules in ALS pathogenesis and identifies Pur-alpha as a novel regulator of SG dynamics.

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“…FUS binds RNA and shuttles between the nucleus and the cytoplasm, participating in nucleo-cytoplasmic RNA shuttling [9]. FUS also plays roles in DNA repair [10–12], transcription [13–23], RNA splicing [22, 24, 25], dendritic RNA transport [26–28] and miRNA biogenesis and function [29, 30]. However, the physiological functions of FUS are still not fully understood.…”

Section: Introductionmentioning

confidence: 99%

Proteomic analysis of FUS interacting proteins provides insights into FUS function and its role in ALS

Kamelgarn

Chen

Kuang

et al. 2016

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease. Mutations in the Fused in Sarcoma/Translocated in Liposarcoma (FUS/TLS) gene cause a subset of familial ALS cases and are also implicated in sporadic ALS. FUS is typically localized to the nucleus. The ALS-related FUS mutations cause cytoplasmic mis-localization and the formation of stress granule-like structures. Abnormal cytoplasmic FUS localization was also found in a subset of frontotemporal dementia (FTLD) cases without FUS mutations. To better understand the function of FUS, we performed wild-type and mutant FUS pull-downs followed by proteomic identification of the interacting proteins. The FUS interacting partners we identified are involved in multiple pathways, including chromosomal organization, transcription, RNA splicing, RNA transport, localized translation, and stress response. FUS interacted with hnRNPA1 and Matrin-3, RNA binding proteins whose mutations were also reported to cause familial ALS, suggesting that hnRNPA1 and Matrin-3 may play common pathogenic roles with FUS. The FUS interactions displayed varied RNA dependence. Numerous FUS interacting partners that we identified are components of exosomes. We found that FUS itself was present in exosomes, suggesting that the secretion of FUS might contribute to the cell-to-cell spreading of FUS pathology. FUS interacting proteins were sequestered into the cytoplasmic mutant FUS inclusions that could lead to their mis-regulation or loss of function, contributing to ALS pathogenesis. Our results provide insights into the physiological functions of FUS as well as important pathways where mutant FUS can interfere with cellular processes and potentially contribute to the pathogenesis of ALS.

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An ALS-associated mutation in the FUS 3′-UTR disrupts a microRNA–FUS regulatory circuitry

Cited by 104 publications

References 31 publications

FUS Mislocalization and Vulnerability to DNA Damage in ALS Patients Derived hiPSCs and Aging Motoneurons

FUS Mislocalization and Vulnerability to DNA Damage in ALS Patients Derived hiPSCs and Aging Motoneurons

Pur-alpha regulates cytoplasmic stress granule dynamics and ameliorates FUS toxicity

Proteomic analysis of FUS interacting proteins provides insights into FUS function and its role in ALS

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