19 results on '"Cestra, G"'
Search Results
2. G4C2 repeat affects nuclear mRNA export pathway in a cellular model of C9orf72-ALS
- Author
-
ROSSI S 1, SERRANO A 1, MIRRA A 2, 3, CESTRA G 4, CARRÌ MT 2, and COZZOLINO M 1
- Subjects
amyotrophic lateral sclerosis ,C9orf72 ,mRNA trafficking ,motor neuron - Abstract
Aims: A non-coding GGGGCC (G4C2) repeat expansion in C9orf72 gene is the most frequent cause of Amyotrophic Lateral Sclerosis (ALS). RNAs containing the repeat, which accumulate as RNA foci in the nucleus and/or cytoplasm of affected cells, bind to various RNA-binding proteins, including several translational regulators, possibly impairing their function. Thus, in order to understand the mechanisms of C9orf72 toxicity, we investigated the functional consequences of G4C2 expression on mRNA trafficking and protein translation. Methods: HeLa cells transfected with (G4C2)31 repeats were used as cellular model. Fluorescence in situ hybridization (FISH), coupled to immunofluorescence analysis, was used to assess RNA foci formation, mRNA distribution and localization of repeat-binding partners. Global protein synthesis was monitored with the SUnSET method. Results: Expression of (G4C2)31 repeats, which induces the formation of nuclear RNA foci, causes stress granules formation, reduction of protein translation and a marked nuclear accumulation of poly-adenylated mRNAs, suggesting that defects in nuclear mRNA export might affect its trafficking and translation. Indeed, C9orf72 repeats interacts with the mRNA export factor NXF1, suggesting a direct interference with NXF1 function. Moreover, overexpression of a dominant negative form of NXF1 reproduces key phenotypes characterizing cells expressing the repeat. Interestingly, modulation of NXF1 expression alters G4C2 RNA foci formation, suggesting a crucial role of this factor in regulating both total mRNA and C9orf72 RNA cellular trafficking. Conclusions: G4C2 expression impairs the NXF1-mediated mRNA export pathway, and the resulting nuclear mRNA retention might affect translation efficiency and contribute to the pathogenesis of C9orf72-ALS.
- Published
- 2016
3. Pur-alpha functionally interacts with FUS carrying ALS-associated mutations
- Author
-
Di Salvio, M, primary, Piccinni, V, additional, Gerbino, V, additional, Mantoni, F, additional, Camerini, S, additional, Lenzi, J, additional, Rosa, A, additional, Chellini, L, additional, Loreni, F, additional, Carrì, M T, additional, Bozzoni, I, additional, Cozzolino, M, additional, and Cestra, G, additional
- Published
- 2015
- Full Text
- View/download PDF
4. An inherited TBX3 alteration in a prenatal case of ulnar-mammary syndrome: Clinical assessment and functional characterization in Drosophila melanogaster.
- Author
-
Bottillo I, D'Alessandro A, Ciccone MP, Cestra G, Di Giacomo G, Silvestri E, Castori M, Brancati F, Lenzi A, Paiardini A, Majore S, Cenci G, and Grammatico P
- Subjects
- Animals, Humans, Female, Upper Extremity Deformities, Congenital genetics, Upper Extremity Deformities, Congenital pathology, Abnormalities, Multiple genetics, Phenotype, Drosophila Proteins genetics, Mutation, Missense genetics, Male, Pregnancy, Mutation genetics, Breast Diseases, T-Box Domain Proteins genetics, Drosophila melanogaster genetics, Ulna abnormalities
- Abstract
Ulnar mammary syndrome (UMS) results from heterozygous variants in the TBX3 gene and impacts limb, tooth, hair, apocrine gland, and genitalia development. The expressivity of UMS is highly variable with no established genotype-phenotype correlations. TBX3 belongs to the Tbx gene family, which encodes transcription factors characterized by the presence of a T-box DNA-binding domain. We describe a fetus exhibiting severe upper limb defects and harboring the novel TBX3:c.400 C > T (p.P134S) variant inherited from the mother who remained clinically misdiagnosed until prenatal diagnosis. Literature revision was conducted to uncover the TBX3 clinical and mutational spectrum. Moreover, we generated a Drosophila humanized model for TBX3 to study the developmental consequences of the p.P134S as well as of other variants targeting different regions of the protein. Phenotypic analysis in flies, coupled with in silico modeling on the TBX3 variants, suggested that the c.400 C > T is UMS-causing and impacts TBX3 localization. Comparative analyses of the fly phenotypes caused by the expression of all variants, demonstrated that missense changes in the T-box domain affect more significantly TBX3 activity than variants outside this domain. To improve the clinicians' recognition of UMS, we estimated the frequency of the main clinical features of the disease. Core features often present pre-pubertally include defects of the ulna and/or of ulnar ray, hypoplastic nipples and/or areolas and, less frequently, genitalia anomalies in young males. These results enhance our understanding of the molecular basis and the clinical spectrum of UMS, shedding light on the functional consequences of TBX3 variants in a developmental context., (© 2024 The Author(s). Journal of Cellular Physiology published by Wiley Periodicals LLC.)
- Published
- 2024
- Full Text
- View/download PDF
5. Cul-4 inhibition rescues spastin levels and reduces defects in hereditary spastic paraplegia models.
- Author
-
Sardina F, Carsetti C, Giorgini L, Fattorini G, Cestra G, and Rinaldo C
- Subjects
- Animals, Humans, Cullin Proteins metabolism, Cullin Proteins genetics, Disease Models, Animal, Drosophila Proteins genetics, Drosophila Proteins metabolism, Haploinsufficiency, Ubiquitination drug effects, Drosophila melanogaster, Spastic Paraplegia, Hereditary genetics, Spastic Paraplegia, Hereditary drug therapy, Spastic Paraplegia, Hereditary metabolism, Spastin metabolism, Spastin genetics
- Abstract
Hereditary spastic paraplegias (HSPs) are degenerative motor neuron diseases characterized by progressive spasticity and weakness in the lower limbs. The most common form of HSP is due to SPG4 gene haploinsufficiency. SPG4 encodes the microtubule severing enzyme spastin. Although, there is no cure for SPG4-HSP, strategies to induce a spastin recovery are emerging as promising therapeutic approaches. Spastin protein levels are regulated by poly-ubiquitination and proteasomal-mediated degradation, in a neddylation-dependent manner. However, the molecular players involved in this regulation are unknown. Here, we show that the Cullin-4-RING E3 ubiquitin ligase complex (CRL4) regulates spastin stability. Inhibition of CRL4 increases spastin levels by preventing its poly-ubiquitination and subsequent degradation in spastin-proficient and in patient derived SPG4 haploinsufficient cells. To evaluate the role of CRL4 complex in spastin regulation in vivo, we developed a Drosophila melanogaster model of SPG4 haploinsufficiency which show alterations of synapse morphology and locomotor activity, recapitulating phenotypical defects observed in patients. Downregulation of the CRL4 complex, highly conserved in Drosophila, rescues spastin levels and the phenotypical defects observed in flies. As a proof of concept of possible pharmacological treatments, we demonstrate a recovery of spastin levels and amelioration of the SPG4-HSP-associated defects both in the fly model and in patient-derived cells by chemical inactivation of the CRL4 complex with NSC1892. Taken together, these findings show that CRL4 contributes to spastin stability regulation and that it is possible to induce spastin recovery and rescue of SPG4-HSP defects by blocking the CRL4-mediated spastin degradation., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Guarantors of Brain.)
- Published
- 2024
- Full Text
- View/download PDF
6. C9orf72 Toxic Species Affect ArfGAP-1 Function.
- Author
-
Rossi S, Di Salvio M, Balì M, De Simone A, Apolloni S, D'Ambrosi N, Arisi I, Cipressa F, Cozzolino M, and Cestra G
- Subjects
- Animals, Humans, Mice, ADP-Ribosylation Factor 1 metabolism, Drosophila genetics, Drosophila metabolism, RNA metabolism, RNA, Messenger genetics, Amyotrophic Lateral Sclerosis metabolism, C9orf72 Protein genetics, C9orf72 Protein metabolism, GTPase-Activating Proteins genetics, GTPase-Activating Proteins metabolism
- Abstract
Compelling evidence indicates that defects in nucleocytoplasmic transport contribute to the pathogenesis of amyotrophic lateral sclerosis (ALS). In particular, hexanucleotide (G4C2) repeat expansions in C9orf72 , the most common cause of genetic ALS, have a widespread impact on the transport machinery that regulates the nucleocytoplasmic distribution of proteins and RNAs. We previously reported that the expression of G4C2 hexanucleotide repeats in cultured human and mouse cells caused a marked accumulation of poly(A) mRNAs in the cell nuclei. To further characterize the process, we set out to systematically identify the specific mRNAs that are altered in their nucleocytoplasmic distribution in the presence of C9orf72 -ALS RNA repeats. Interestingly, pathway analysis showed that the mRNAs involved in membrane trafficking are particularly enriched among the identified mRNAs. Most importantly, functional studies in cultured cells and Drosophila indicated that C9orf72 toxic species affect the membrane trafficking route regulated by ADP-Ribosylation Factor 1 GTPase Activating Protein (ArfGAP-1), which exerts its GTPase-activating function on the small GTPase ADP-ribosylation factor 1 to dissociate coat proteins from Golgi-derived vesicles. We demonstrate that the function of ArfGAP-1 is specifically affected by expanded C9orf72 RNA repeats, as well as by C9orf72 -related dipeptide repeat proteins (C9-DPRs), indicating the retrograde Golgi-to-ER vesicle-mediated transport as a target of C9orf72 toxicity.
- Published
- 2023
- Full Text
- View/download PDF
7. Biallelic mutations in RNF220 cause laminopathies featuring leukodystrophy, ataxia and deafness.
- Author
-
Sferra A, Fortugno P, Motta M, Aiello C, Petrini S, Ciolfi A, Cipressa F, Moroni I, Leuzzi V, Pieroni L, Marini F, Boespflug Tanguy O, Eymard-Pierre E, Danti FR, Compagnucci C, Zambruno G, Brusco A, Santorelli FM, Chiapparini L, Francalanci P, Loizzo AL, Tartaglia M, Cestra G, and Bertini E
- Subjects
- Adolescent, Amino Acid Sequence, Animals, Ataxia diagnosis, COS Cells, Child, Chlorocebus aethiops, Deafness diagnosis, Drosophila, Female, HEK293 Cells, Humans, Laminopathies diagnosis, Male, Pedigree, Young Adult, Alleles, Ataxia genetics, Deafness genetics, Laminopathies genetics, Mutation genetics, Ubiquitin-Protein Ligases genetics
- Abstract
Leukodystrophies are a heterogeneous group of rare inherited disorders that mostly involve the white matter of the CNS. These conditions are characterized by primary glial cell and myelin sheath pathology of variable aetiology, which causes secondary axonal degeneration, generally emerging with disease progression. Whole exome sequencing performed in five large consanguineous nuclear families allowed us to identify homozygosity for two recurrent missense variants affecting highly conserved residues of RNF220 as the causative event underlying a novel form of leukodystrophy with ataxia and sensorineural deafness. We report these two homozygous missense variants (p.R363Q and p.R365Q) in the ubiquitin E3 ligase RNF220 as the underlying cause of this novel form of leukodystrophy with ataxia and sensorineural deafness that includes fibrotic cardiomyopathy and hepatopathy as associated features in seven consanguineous families. Mass spectrometry analysis identified lamin B1 as the RNF220 binding protein and co-immunoprecipitation experiments demonstrated reduced binding of both RNF220 mutants to lamin B1. We demonstrate that RNF220 silencing in Drosophila melanogaster specifically affects proper localization of lamin Dm0, the fly lamin B1 orthologue, promotes its aggregation and causes a neurodegenerative phenotype, strongly supporting the functional link between RNF220 and lamin B1. Finally, we demonstrate that RNF220 plays a crucial role in the maintenance of nuclear morphology; mutations in primary skin fibroblasts determine nuclear abnormalities such as blebs, herniations and invaginations, which are typically observed in cells of patients affected by laminopathies. Overall, our data identify RNF220 as a gene implicated in leukodystrophy with ataxia and sensorineural deafness and document a critical role of RNF220 in the regulation of nuclear lamina. Our findings provide further evidence on the direct link between nuclear lamina dysfunction and neurodegeneration., (© The Author(s) (2021). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2021
- Full Text
- View/download PDF
8. Chemical chaperones targeted to the endoplasmic reticulum (ER) and lysosome prevented neurodegeneration in a C9orf72 repeat expansion drosophila amyotrophic lateral sclerosis (ALS) model.
- Author
-
Azoulay-Ginsburg S, Di Salvio M, Weitman M, Afri M, Ribeiro S, Ebbinghaus S, Cestra G, and Gruzman A
- Subjects
- Amyotrophic Lateral Sclerosis genetics, Amyotrophic Lateral Sclerosis physiopathology, Animals, DNA Repeat Expansion genetics, Disease Models, Animal, Drosophila melanogaster, Endoplasmic Reticulum drug effects, Lysosomes metabolism, Magnetic Resonance Imaging, Molecular Chaperones chemical synthesis, Molecular Chaperones chemistry, Phenylbutyrates chemical synthesis, Phenylbutyrates chemistry, Amyotrophic Lateral Sclerosis drug therapy, C9orf72 Protein genetics, Molecular Chaperones pharmacology, Phenylbutyrates pharmacology
- Abstract
Background: ALS is an incurable neuromuscular degenerative disorder. A familiar form of the disease (fALS) is related to point mutations. The most common one is an expansion of a noncoding GGGGCC hexanucleotide repeat of the C9orf72 gene on chromosome 9p21. An abnormal translation of the C9orf72 gene generates dipeptide repeat proteins that aggregate in the brain. One of the classical approaches for developing treatment against protein aggregation-related diseases is to use chemical chaperones (CSs). In this work, we describe the development of novel 4-phenylbutyric acid (4-PBA) lysosome/ER-targeted derivatives. We assumed that 4-PBA targeting to specific organelles, where protein degradation takes place, might reduce the 4-PBA effective concentration., Methods: Organic chemistry synthetic methods and solid-phase peptide synthesis (SPPS) were used for preparing the 4-PBA derivatives. The obtained compounds were evaluated in an ALS Drosophila model that expressed C9orf72 repeat expansion, causing eye degeneration. Targeting to lysosome was validated by the
19 F-nuclear magnetic resonance (NMR) technique., Results: Several synthesized compounds exhibited a significant biological effect by ameliorating the eye degeneration. They blocked the neurodegeneration of fly retina at different efficacy levels. The most active CS was compound 9, which is a peptide derivative and was targeted to ER. Another active compound targeted to lysosome was compound 4., Conclusions: Novel CSs were more effective than 4-PBA; therefore, they might be used as a new class of drug candidates to treat ALS and other protein misfolding disorders.- Published
- 2021
- Full Text
- View/download PDF
9. HDAC1 inhibition ameliorates TDP-43-induced cell death in vitro and in vivo.
- Author
-
Sanna S, Esposito S, Masala A, Sini P, Nieddu G, Galioto M, Fais M, Iaccarino C, Cestra G, and Crosio C
- Subjects
- Amyotrophic Lateral Sclerosis genetics, Animals, Cell Death physiology, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Frontotemporal Lobar Degeneration metabolism, Frontotemporal Lobar Degeneration pathology, Histone Deacetylase 1 genetics, Humans, Inclusion Bodies metabolism, Mice, Mutation genetics, Amyotrophic Lateral Sclerosis pathology, Cell Death drug effects, DNA-Binding Proteins pharmacology, Histone Deacetylase 1 metabolism
- Abstract
TDP-43 pathology is a disease hallmark that characterizes both amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD-TDP). TDP-43 undergoes several posttranslational modifications that can change its biological activities and its aggregative propensity, which is a common hallmark of different neurodegenerative conditions. New evidence is provided by the current study pointing at TDP-43 acetylation in ALS cellular models. Using both in vitro and in vivo approaches, we demonstrate that TDP-43 interacts with histone deacetylase 1 (HDAC1) via RRM1 and RRM2 domains, that are known to contain the two major TDP-43 acetylation sites, K142 and K192. Moreover, we show that TDP-43 is a direct transcriptional activator of CHOP promoter and this activity is regulated by acetylation. Finally and most importantly, we observe both in cell culture and in Drosophila that a HDCA1 reduced level (genomic inactivation or siRNA) or treatment with pan-HDAC inhibitors exert a protective role against WT or pathological mutant TDP-43 toxicity, suggesting TDP-43 acetylation as a new potential therapeutic target. HDAC inhibition efficacy in neurodegeneration has long been debated, but future investigations are warranted in this area. Selection of more specific HDAC inhibitors is still a promising option for neuronal protection especially as HDAC1 appears as a downstream target of both TDP- 43 and FUS, another ALS-related gene.
- Published
- 2020
- Full Text
- View/download PDF
10. UsnRNP trafficking is regulated by stress granules and compromised by mutant ALS proteins.
- Author
-
Rossi S, Rompietti V, Antonucci Y, Giovannini D, Scopa C, Scaricamazza S, Scardigli R, Cestra G, Serafino A, Carrì MT, D'Ambrosi N, and Cozzolino M
- Subjects
- Alternative Splicing, Amyotrophic Lateral Sclerosis metabolism, Animals, C9orf72 Protein genetics, Cell Nucleus genetics, Cytoplasm genetics, DNA-Binding Proteins genetics, Humans, Mice, Motor Neurons pathology, Mutation, Protein Transport genetics, RNA-Binding Protein FUS genetics, Amyotrophic Lateral Sclerosis genetics, Mutant Proteins genetics, Ribonucleoproteins, Small Nuclear genetics
- Abstract
Activation of the integrated stress response (ISR), alterations in nucleo-cytoplasmic (N/C) transport and changes in alternative splicing regulation are all common traits of the pathogenesis of Amyotrophic Lateral Sclerosis (ALS). However, whether these processes act independently from each other, or are part of a coordinated mechanism of gene expression regulation that is affected in pathogenic conditions, is still rather undefined. To answer these questions, in this work we set out to characterise the functional connections existing between ISR activation and nucleo-cytosol trafficking and nuclear localization of spliceosomal U-rich small nuclear ribonucleoproteins (UsnRNPs), the core constituents of the spliceosome, and to study how ALS-linked mutant proteins affect this interplay. Activation of the ISR induces a profound reorganization of nuclear Gems and Cajal bodies, the membrane-less particles that assist UsnRNP maturation and storage. This effect requires the cytoplasmic assembly of SGs and is associated to the disturbance of the nuclear import of UsnRNPs by the snurportin-1/importin-β1 system. Notably, these effects are reversed by both inhibiting the ISR or upregulating importin-β1. This indicates that SGs are major determinants of Cajal bodies assembly and that the modulation of N/C trafficking of UsnRNPs might control alternative splicing in response to stress. Importantly, the dismantling of nuclear Gems and Cajal bodies by ALS-linked mutant FUS or C9orf72-derived dipeptide repeat proteins is halted by overexpression of importin-β1, but not by inhibition of the ISR. This suggests that changes in the nuclear localization of the UsnRNP complexes induced by mutant ALS proteins are uncoupled from ISR activation, and that defects in the N/C trafficking of UsnRNPs might play a role in ALS pathogenesis., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
11. TUBB Variants Underlying Different Phenotypes Result in Altered Vesicle Trafficking and Microtubule Dynamics.
- Author
-
Sferra A, Petrini S, Bellacchio E, Nicita F, Scibelli F, Dentici ML, Alfieri P, Cestra G, Bertini ES, and Zanni G
- Subjects
- Amino Acid Sequence, Cell Movement drug effects, Child, DNA Mutational Analysis, Epidermal Growth Factor metabolism, Fibroblasts drug effects, Fibroblasts metabolism, Humans, Magnetic Resonance Imaging, Models, Molecular, Nocodazole pharmacology, Phenotype, Protein Transport, Transferrin metabolism, Tubulin chemistry, Microtubules metabolism, Mutation genetics, Transport Vesicles metabolism, Tubulin genetics
- Abstract
Tubulinopathies are rare neurological disorders caused by alterations in tubulin structure and function, giving rise to a wide range of brain abnormalities involving neuronal proliferation, migration, differentiation and axon guidance. TUBB is one of the ten β-tubulin encoding genes present in the human genome and is broadly expressed in the developing central nervous system and the skin. Mutations in TUBB are responsible for two distinct pathological conditions: the first is characterized by microcephaly and complex structural brain malformations and the second, also known as "circumferential skin creases Kunze type" (CSC-KT), is associated to neurological features, excess skin folding and growth retardation. We used a combination of immunocytochemical and cellular approaches to explore, on patients' derived fibroblasts, the functional consequences of two TUBB variants: the novel mutation (p.N52S), associated with basal ganglia and cerebellar dysgenesis, and the previously reported variant (p.M73T), linked to microcephaly, corpus callosum agenesis and CSC-KT skin phenotype. Our results demonstrate that these variants impair microtubule (MT) function and dynamics. Most importantly, our studies show an altered epidermal growth factor (EGF) and transferrin (Tf) intracellular vesicle trafficking in both patients' fibroblasts, suggesting a specific role of TUBB in MT-dependent vesicular transport., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.
- Published
- 2020
- Full Text
- View/download PDF
12. A Lipophilic 4-Phenylbutyric Acid Derivative That Prevents Aggregation and Retention of Misfolded Proteins.
- Author
-
Azoulay-Ginsburg S, Trobiani L, Setini A, Favaloro FL, Giorda E, Jacob A, Hauschner H, Levy L, Cestra G, De Jaco A, and Gruzman A
- Subjects
- Animals, Cell Adhesion Molecules, Neuronal chemistry, Cell Adhesion Molecules, Neuronal genetics, Cell Adhesion Molecules, Neuronal metabolism, Cell Survival drug effects, HEK293 Cells, Humans, Membrane Proteins chemistry, Membrane Proteins genetics, Membrane Proteins metabolism, Mutagenesis, Site-Directed, Nerve Tissue Proteins chemistry, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism, PC12 Cells, Phenylbutyrates pharmacology, Protein Aggregates drug effects, Protein Folding, Proteins metabolism, Rats, Phenylbutyrates chemistry, Proteins chemistry
- Abstract
Chemical chaperones prevent protein aggregation. However, the use of chemical chaperones as drugs against diseases due to protein aggregation is limited by the very high active concentrations (mm range) required to mediate their effect. One of the most common chemical chaperones is 4-phenylbutyric acid (4-PBA). Despite its unfavorable pharmacokinetic properties, 4-PBA was approved as a drug to treat ornithine cycle diseases. Here, we report that 2-isopropyl-4-phenylbutanoic acid (5) has been found to be 2-10-fold more effective than 4-PBA in several in vitro models of protein aggregation. Importantly, compound 5 reduced the secretion rate of autism-linked Arg451Cys Neuroligin3 (R451C NLGN3)., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2020
- Full Text
- View/download PDF
13. Recessive mutations in the neuronal isoforms of DST, encoding dystonin, lead to abnormal actin cytoskeleton organization and HSAN type VI.
- Author
-
Fortugno P, Angelucci F, Cestra G, Camerota L, Ferraro AS, Cordisco S, Uccioli L, Castiglia D, De Angelis B, Kurth I, Kornak U, and Brancati F
- Subjects
- Actins metabolism, Adult, Aged, Amino Acid Sequence, Animals, COS Cells, Cell Adhesion, Cell Movement, Chlorocebus aethiops, Dermis pathology, Dystonin chemistry, Family, Female, Fibroblasts metabolism, Fibroblasts pathology, HEK293 Cells, Humans, Male, Middle Aged, Protein Binding, Protein Isoforms genetics, Actin Cytoskeleton pathology, Dystonin genetics, Genes, Recessive, Hereditary Sensory and Autonomic Neuropathies genetics, Mutation genetics, Neurons metabolism
- Abstract
Hereditary sensory and autonomic neuropathies (HSAN) are clinically and genetically heterogeneous disorders, characterized by a progressive sensory neuropathy often complicated by ulcers and amputations, with variable motor and autonomic involvement. Several pathways have been implicated in the pathogenesis of neuronal degeneration in HSAN, while recent observations point to an emerging role of cytoskeleton organization and function. Here, we report novel biallelic mutations in the DST gene encoding dystonin, a large cytolinker protein of the plakin family, in an adult form of HSAN type VI. Affected individuals harbored the premature termination codon variant p.(Lys4330*) in trans with the p.(Ala203Glu) change affecting a highly conserved residue in an isoform-specific N-terminal region of dystonin. Functional studies showed defects in actin cytoskeleton organization and consequent delayed cell adhesion, spreading and migration, while recombinant p.Ala203Glu dystonin loses the ability to bind actin. Our data aid in the clinical and molecular delineation of HSAN-VI and suggest a central role for cell-motility and cytoskeletal defects in its pathogenesis possibly interfering with the neuronal outgrowth and guidance processes., (© 2018 Wiley Periodicals, Inc.)
- Published
- 2019
- Full Text
- View/download PDF
14. Publisher Correction: Functional interaction between FUS and SMN underlies SMA-like splicing changes in wild-type hFUS mice.
- Author
-
Mirra A, Rossi S, Scaricamazza S, Salvio MD, Salvatori I, Valle C, Rusmini P, Poletti A, Cestra G, Carrì MT, and Cozzolino M
- Abstract
A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.
- Published
- 2018
- Full Text
- View/download PDF
15. Functional interaction between FUS and SMN underlies SMA-like splicing changes in wild-type hFUS mice.
- Author
-
Mirra A, Rossi S, Scaricamazza S, Di Salvio M, Salvatori I, Valle C, Rusmini P, Poletti A, Cestra G, Carrì MT, and Cozzolino M
- Abstract
Several of the identified genetic factors in Amyotrophic Lateral Sclerosis (ALS) point to dysfunction in RNA processing as a major pathogenic mechanism. However, whether a precise RNA pathway is particularly affected remains unknown. Evidence suggests that FUS, that is mutated in familial ALS, and SMN, the causative factor in Spinal Muscular Atrophy (SMA), cooperate to the same molecular pathway, i.e. regulation of alternative splicing, and that disturbances in SMN-regulated functions, either caused by depletion of SMN protein (as in the case of SMA) or by pathogenic interactions between FUS and SMN (as in the case of ALS) might be a common theme in both diseases. In this work, we followed these leads and tested their pathogenic relevance in vivo. FUS-associated ALS recapitulates, in transgenic mice, crucial molecular features that characterise mouse models of SMA, including defects in snRNPs distribution and in the alternative splicing of genes important for motor neurons. Notably, altering SMN levels by haploinsufficiency or overexpression does not impact the phenotypes of mouse or Drosophila models of FUS-mediated toxicity. Overall, these findings suggest that FUS and SMN functionally interact and that FUS may act downstream of SMN-regulated snRNP assembly in the regulation of alternative splicing and gene expression.
- Published
- 2017
- Full Text
- View/download PDF
16. Control of mRNA Translation in ALS Proteinopathy.
- Author
-
Cestra G, Rossi S, Di Salvio M, and Cozzolino M
- Abstract
Cells robustly reprogram gene expression during stress generated by protein misfolding and aggregation. In this condition, cells assemble the bulk of mRNAs into translationally silent stress granules (SGs), while they sustain the translation of specific mRNAs coding for proteins that are needed to overcome cellular stress. Alterations of this process are deeply associated to neurodegeneration. This is the case of amyotrophic lateral sclerosis (ALS), a neurodegenerative disorder caused by a selective loss of motor neurons. Indeed, impairment of protein homeostasis as well as alterations of RNA metabolism are now recognized as major players in the pathogenesis of ALS. In particular, evidence shows that defective mRNA transport and translation are implicated. Here, we provide a review of what is currently known about altered mRNA translation in ALS and how this impacts on the ability of affected cells to cope with proteotoxic stress.
- Published
- 2017
- Full Text
- View/download PDF
17. nArgBP2 regulates excitatory synapse formation by controlling dendritic spine morphology.
- Author
-
Lee SE, Kim Y, Han JK, Park H, Lee U, Na M, Jeong S, Chung C, Cestra G, and Chang S
- Subjects
- Adaptor Proteins, Signal Transducing genetics, Animals, Bipolar Disorder genetics, Bipolar Disorder metabolism, Gene Knockdown Techniques, Mice, Protein Isoforms genetics, Protein Isoforms metabolism, Rats, Rats, Sprague-Dawley, Synapses genetics, Adaptor Proteins, Signal Transducing metabolism, Dendritic Spines metabolism, Synapses metabolism
- Abstract
Neural Abelson-related gene-binding protein 2 (nArgBP2) was originally identified as a protein that directly interacts with synapse-associated protein 90/postsynaptic density protein 95-associated protein 3 (SAPAP3), a postsynaptic scaffolding protein critical for the assembly of glutamatergic synapses. Although genetic deletion of nArgBP2 in mice leads to manic/bipolar-like behaviors resembling many aspects of symptoms in patients with bipolar disorder, the actual function of nArgBP2 at the synapse is completely unknown. Here, we found that the knockdown (KD) of nArgBP2 by specific small hairpin RNAs (shRNAs) resulted in a dramatic change in dendritic spine morphology. Reintroducing shRNA-resistant nArgBP2 reversed these defects. In particular, nArgBP2 KD impaired spine-synapse formation such that excitatory synapses terminated mostly at dendritic shafts instead of spine heads in spiny neurons, although inhibitory synapse formation was not affected. nArgBP2 KD further caused a marked increase of actin cytoskeleton dynamics in spines, which was associated with increased Wiskott-Aldrich syndrome protein-family verprolin homologous protein 1 (WAVE1)/p21-activated kinase (PAK) phosphorylation and reduced activity of cofilin. These effects of nArgBP2 KD in spines were rescued by inhibiting PAK or activating cofilin combined with sequestration of WAVE. Together, our results suggest that nArgBP2 functions to regulate spine morphogenesis and subsequent spine-synapse formation at glutamatergic synapses. They also raise the possibility that the aberrant regulation of synaptic actin filaments caused by reduced nArgBP2 expression may contribute to the manifestation of the synaptic dysfunction observed in manic/bipolar disorder.
- Published
- 2016
- Full Text
- View/download PDF
18. Translational repression in the pathogenesis of FUS- and C9orf72-dependent ALS.
- Author
-
Cozzolino M, Rossi S, and Cestra G
- Published
- 2015
- Full Text
- View/download PDF
19. Nuclear accumulation of mRNAs underlies G4C2-repeat-induced translational repression in a cellular model of C9orf72 ALS.
- Author
-
Rossi S, Serrano A, Gerbino V, Giorgi A, Di Francesco L, Nencini M, Bozzo F, Schininà ME, Bagni C, Cestra G, Carrì MT, Achsel T, and Cozzolino M
- Subjects
- Amyotrophic Lateral Sclerosis pathology, Animals, C9orf72 Protein, DNA-Binding Proteins, Eukaryotic Initiation Factor-2 metabolism, Fragile X Mental Retardation Protein metabolism, HeLa Cells, Humans, Intracellular Space metabolism, Mice, Motor Neurons metabolism, Phosphorylation, Poly(A)-Binding Proteins metabolism, Protein Binding, RNA Splicing genetics, RNA, Messenger genetics, RNA, Messenger metabolism, Transcription Factors, Amyotrophic Lateral Sclerosis metabolism, Cell Nucleus metabolism, Models, Biological, Protein Biosynthesis, Proteins metabolism, Trinucleotide Repeat Expansion
- Abstract
A common feature of non-coding repeat expansion disorders is the accumulation of RNA repeats as RNA foci in the nucleus and/or cytoplasm of affected cells. These RNA foci can be toxic because they sequester RNA-binding proteins, thus affecting various steps of post-transcriptional gene regulation. However, the precise step that is affected by C9orf72 GGGGCC (G4C2) repeat expansion, the major genetic cause of amyotrophic lateral sclerosis (ALS), is still poorly defined. In this work, we set out to characterise these mechanisms by identifying proteins that bind to C9orf72 RNA. Sequestration of some of these factors into RNA foci was observed when a (G4C2)31 repeat was expressed in NSC34 and HeLa cells. Most notably, (G4C2)31 repeats widely affected the distribution of Pur-alpha and its binding partner fragile X mental retardation protein 1 (FMRP, also known as FMR1), which accumulate in intra-cytosolic granules that are positive for stress granules markers. Accordingly, translational repression is induced. Interestingly, this effect is associated with a marked accumulation of poly(A) mRNAs in cell nuclei. Thus, defective trafficking of mRNA, as a consequence of impaired nuclear mRNA export, might affect translation efficiency and contribute to the pathogenesis of C9orf72 ALS., (© 2015. Published by The Company of Biologists Ltd.)
- Published
- 2015
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.