ALS-linked FUS exerts a gain of toxic function involving aberrant p38 MAPK activation

Sama, Reddy Ranjith Kumar; Fallini, Claudia; Gatto, Rodolfo G.; McKeon, Jeanne E.; Song, Yuyu; Rotunno, Melissa; Penaranda, Saul; Abdurakhmanov, Izrail; Landers, John E.; Morfini, Gerardo; Brady, Scott T.; Bosco, Daryl A.

doi:10.1038/s41598-017-00091-1

Cited by 48 publications

(54 citation statements)

References 69 publications

(133 reference statements)

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“…MAPKs and GSK3 are implicated in proper SG function and disease. For instance, aberrant MAPK p38 activity induces a gain of function for the ALS-linked SG component FUS (65), and, conversely, inhibition of MAPK signaling improves survival of SOD1 E100G mutant motor neurons derived from ALS patient induced pluripotent stem cells (66). Likewise, tight regulation of GSK3 activity appears to underlie proper SG responses, given that GSK3 inhibition dramatically reduces stress granule formation (12,67), and aberrant activation of GSK3 is associated with ALS (68).…”

Section: Phosphorylation Of Gle1a Regulates Ddx3 and Stress Responsementioning

confidence: 99%

MAPK- and glycogen synthase kinase 3–mediated phosphorylation regulates the DEAD-box protein modulator Gle1 for control of stress granule dynamics

Aditi

Mason²,

Sharma³

et al. 2019

Journal of Biological Chemistry

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Rapid expression of critical stress response factors is a key survival strategy for diseased or stressed cells. During cell stress, translation is inhibited, and a pre-existing pool of cytoplasmic mRNA-protein complexes reversibly assembles into cytoplasmic stress granules (SGs). Gle1 is a conserved modulator of RNA-dependent DEAD-box proteins required for mRNA export, translation, and stress responses. Proper Gle1 function is critical as reflected by some human diseases such as developmental and neurodegenerative disorders and some cancers linked to gle1 mutations. However, the mechanism by which Gle1 controls SG formation is incompletely understood. Here, we show that human Gle1 is regulated by phosphorylation during heat shock stress. In HeLa cells, stress-induced Gle1 hyperphosphorylation was dynamic, primarily in the cytoplasmic pool, and followed changes in translation factors. MS analysis identified 14 phosphorylation sites in the Gle1A isoform, six of which clustered in an intrinsically disordered, low-complexity N-terminal region flanking the coil-coiled self-association domain. Of note, two mitogen-activated protein kinases (MAPKs), extracellular signal-regulated kinase (ERK) and

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Section: Phosphorylation Of Gle1a Regulates Ddx3 and Stress Responsementioning

confidence: 99%

MAPK- and glycogen synthase kinase 3–mediated phosphorylation regulates the DEAD-box protein modulator Gle1 for control of stress granule dynamics

Aditi

Mason²,

Sharma³

et al. 2019

Journal of Biological Chemistry

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“…Disruptions in axonal transport have been linked to the M337V TARDBP mutation in a range of in vitro and Drosophila larval models (Wang et al 2013;Alami et al 2014;Baldwin et al 2016), and, while the severe, frameshift FUS mutation modelled in Fus Δ14/+ mice has not previously been assessed, transport perturbations have been reported in several mutant FUS models, including Drosophila larvae (Baldwin et al 2016), isolated squid axoplasm (Sama et al 2017), and human motor neurons derived from induced pluripotent stem cells (iPSCs) (Guo et al 2017). Why then do Fus Δ14/+ mice not show impaired signalling endosome transport at least until a very late disease stage?…”

Section: Discussionmentioning

confidence: 99%

ALS mice carrying pathological mutant TDP-43, but not mutant FUS, display axonal transport defects in vivo

Sleigh

Tosolini

Gordon

et al. 2018

Preprint

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Amyotrophic lateral sclerosis (ALS) is a fatal, progressive neurodegenerative disease resulting from a complex interplay between genetics and environment. Impairments in the basic neuronal process of axonal transport have been identified in several ALS models. However, in vivo evidence of early/pre-symptomatic deficiencies in neuronal cargo trafficking remains limited, thus the pathogenic importance of axonal transport to the ALS disease spectrum remains to be fully resolved. We therefore analysed the in vivo dynamics of retrogradely transported, neurotrophin-containing signalling endosomes in motor neuron axons of two new mouse models of ALS that have mutations in different RNA processing genes (Tardbp and Fus). TDP-43 M337V mice, which show neuromuscular pathology but no overt motor neuron loss, displayed in vivo perturbations in axonal transport that manifested between 1.5 and 3 months and preceded motor symptom onset. In contrast, signalling endosome transport remained largely unaffected in mutant Fus Δ14/+ mice, despite 20% motor neuron loss. These findings indicate that deficiencies in retrograde neurotrophin signalling and axonal transport are not common to all ALS-linked genes, and that there are inherent and mechanistic distinctions in the pathogenesis of ALS caused by mutations in different RNA processing genes.

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“…Further, the effects of ALS-linked FUS on calcium-mediated motor neuron toxicity is exacerbated by expression of the mutant protein in astrocytes 51,8 . Most ALS-linked FUS mutations are located within the NLS 29 and induce a shift in the nucleocytoplasmic equilibrium of the protein toward the cytoplasm, where it is believed to exert a gain of toxic function 26,68 (Fig. 3).…”

Section: Discussionmentioning

confidence: 99%

“…Primary motor neurons were isolated from embryonic day 12.5 murine spinal cords as described 68 . Briefly, after dissociation in 0.1% trypsin (Worthington LS003707, Columbus, OH, USA) at 37°C for 12 minutes, primary motor neurons were purified using a 6% Optiprep (MilliporeSigma D1556) density gradient and plated on glass coverslips coated with 0.5g/L poly-ornithine and natural mouse laminin (Thermo Fisher 23017015, Waltham, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Cortical neuron immunofluorescence experiments were conducted as described 12,26 using antibodies listed in Supplementary Table 1 . Primary motor neuron samples were blocked in 4% bovine serum albumin for 45 minutes and hybridized overnight at 4°C with primary antibodies ( Supplementary Table 1 ) and AlexaFluor-conjugated secondary antibodies 68 .…”

Section: Methodsmentioning

confidence: 99%

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FUS/TLS undergoes calcium-mediated nuclear egress during excitotoxic stress and is required for Gria2 mRNA processing

Tischbein

Yan

et al. 2018

Preprint

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27Excitotoxic levels of glutamate represent a physiological stress that is strongly linked to 28 amyotrophic lateral sclerosis (ALS) and other neurological disorders. Emerging evidence 29 indicates a role for neurodegenerative disease linked RNA-binding proteins (RBPs) in the cellular 30 stress response. However, the relationships between excitotoxicity, RBP function and pathology 31 have not been explored. Here, we found that excitotoxicity induced the translocation of select 32 ALS-linked RBPs from the nucleus to the cytoplasm within neurons. RBPs affected by 33 excitotoxicity include TAR DNA-binding protein 43 (TDP-43) and, most robustly, fused in 34 sarcoma/translocated in liposarcoma (FUS/TLS). FUS translocation occurs through a calcium-35 dependent mechanism and coincides with striking alterations in nucleocytoplasmic transport.36 Further, glutamate-induced upregulation of Gria2 in neurons was dependent on FUS expression, 37 consistent with a functional role for FUS under excitotoxic stress. These findings reveal a link 38 between prominent factors in neurodegenerative disease, namely excitotoxicity, disease-39 associated RBPs and nucleocytoplasmic transport.40 41 on poly-ornithine (final concentration of 1.5 µg/mL; MilliporeSigma P4957) coated plates or 104 coverslips. Neuronal cultures were grown under standard culture conditions (37°C, 5% CO2/95% 105 air) fed every 3-4 days by adding half volumes of supplemented neurobasal media to each 106 well/dish, with additional half changes of media occurring every other feeding. Unless indicated, 107 during the first feeding (DIV 2 or 3) neuron cultures were also treated with a final concentration of 108 0.5-1µM Cytosine β-D-arabinofuranoside hydrochloride (MilliporeSigma C6645) to inhibit non-109 neuronal cell growth. Experiments were performed on day in vitro (DIV) 14-16. 110Primary motor neurons were isolated from embryonic day 12.5 murine spinal cords as 111 described 68 . Briefly, after dissociation in 0.1% trypsin (Worthington LS003707, Columbus, OH, 112 USA) at 37°C for 12 minutes, primary motor neurons were purified using a 6% Optiprep 113 (MilliporeSigma D1556) density gradient and plated on glass coverslips coated with 0.5g/L poly-

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ALS-linked FUS exerts a gain of toxic function involving aberrant p38 MAPK activation

Cited by 48 publications

References 69 publications

MAPK- and glycogen synthase kinase 3–mediated phosphorylation regulates the DEAD-box protein modulator Gle1 for control of stress granule dynamics

MAPK- and glycogen synthase kinase 3–mediated phosphorylation regulates the DEAD-box protein modulator Gle1 for control of stress granule dynamics

ALS mice carrying pathological mutant TDP-43, but not mutant FUS, display axonal transport defects in vivo

FUS/TLS undergoes calcium-mediated nuclear egress during excitotoxic stress and is required for Gria2 mRNA processing

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