The 14-3-3/spastin pathway coordinates neurite regeneration and recovery of spinal cord injury by targeting microtubule dynamics DOI Open Access
Qiuling Liu, Hua Yang, Jianxian Luo

et al.

Published: Nov. 14, 2023

Axon regeneration is abortive in the central nervous system following injury. Orchestrating microtubule dynamics has emerged as a promising approach to improve axonal regeneration. The severing enzyme spastin essential for development and through remodeling of arrangement. To date, however, little known regarding mechanisms underlying action neural after spinal cord Here, we use glutathione transferase pulldown immunoprecipitation assays demonstrate that 14-3-3 interacts with spastin, both vivo vitro, via Ser233 phosphorylation. Moreover, show protects from degradation by inhibiting ubiquitination pathway upregulates spastin-dependent ability. Furthermore, improving interaction between Fusicoccin (FC-A) promotes neurite outgrowth vitro . Western blot immunofluorescence results revealed protein upregulated neuronal compartment injury In addition, administration FC-A not only locomotor recovery, but also nerve contusion lateral hemisection models. However, application inhibitor spastazoline successfully reverses these phenomena. Taken together, indicate molecular switch regulates levels, 14-3-3/spastin an important target regulation

Language: Английский

Doublecortin restricts neuronal branching by regulating tubulin polyglutamylation DOI Creative Commons
Muriel Sébastien, Alexandra Paquette, Emily N. P. Prowse

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Feb. 18, 2025

Doublecortin is a neuronal microtubule-associated protein that regulates microtubule structure in neurons. Mutations cause lissencephaly and subcortical band heterotopia by impairing migration. We use CRISPR/Cas9 to knock-out the gene induced pluripotent stem cells differentiate into cortical DCX-KO neurons show reduced velocities of nuclear movements an increased number neurites early development, consistent with previous findings. Neurite branching regulated host proteins, as well polymerization dynamics. However, EB comet dynamics are unchanged Rather, we observe significant reduction α-tubulin polyglutamylation Polyglutamylation levels rescued expression or TTLL11, glutamylase. Using U2OS orthogonal model system, DCX TTLL11 act synergistically promote polyglutamylation. propose acts positive regulator restricts neurite branching. Our results indicate unexpected role for homeostasis tubulin code. Lissencephaly severe neurodevelopmental disease often caused mutations Dcx gene. human cellular lissencephaly, authors report activating

Language: Английский

Citations

1
Cul-4 inhibition rescues spastin levels and reduces defects in hereditary spastic paraplegia models DOI Creative Commons
Francesca Sardina, Claudia Carsetti, Ludovica Giorgini

et al.

Brain, Journal Year: 2024, Volume and Issue: unknown

Published: March 29, 2024

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. encodes microtubule severing enzyme spastin. Although, there no cure for SPG4-HSP, strategies induce a spastin recovery emerging as promising therapeutic approaches. Spastin protein levels regulated poly-ubiquitination proteasomal-mediated degradation, neddylation-dependent manner. However, molecular players involved this regulation unknown. Here, we show that Cullin-4-Ring E3 ubiquitin ligase complex (CRL4) regulates stability. Inhibition CRL4 increases preventing its subsequent degradation spastin-proficient patient derived haploinsufficient cells. To evaluate role vivo, developed Drosophila melanogaster model haploinsufficiency which alterations synapse morphology locomotor activity, recapitulating phenotypical defects observed patients. Downregulation complex, highly conserved Drosophila, rescues flies. As proof concept possible pharmacological treatments, demonstrate amelioration SPG4-HSP-associated both fly patient-derived cells chemical inactivation with NSC1892. Taken together, these findings contributes stability it rescue SPG4-HSP blocking CRL4-mediated degradation.

Language: Английский

Citations

6
Targeting MDM2 affects spastin protein levels and functions: implications for HSP treatment DOI Creative Commons
Francesca Sardina, Federica Polverino,

Sonia Valentini

et al.

Cell Death Discovery, Journal Year: 2025, Volume and Issue: 11(1)

Published: Feb. 7, 2025

Spastin is a microtubule (MT) severing enzyme that regulates several cell functions associated with MT dynamics. A reduction in spastin protein levels responsible for approximately 40% of cases Hereditary Spastic Paraplegia (HSP), neurodegenerative disease. Currently, there no cure HSP but strategies to induce recovery are emerging as potential therapeutic approaches. Here, we show MDM2 interacts MT-interacting and trafficking (MIT) domain. By biochemical functional experiments, demonstrate binds its post-transcriptional manner independently the E3 ubiquitin ligase activity. Of relevance, treatment spastin-deficient cells inhibitor Nutlin-3a can restore functions, such cytokinetic abscission sorting transferrin receptor. These findings identify novel interactor druggable regulator levels.

Language: Английский

Citations

0
Choroid plexuses carry nodal-like cilia that undergo axoneme regression from early adult stage DOI Creative Commons

Kim Hoa Ho,

Adrien Candat,

Valentina Scarpetta

et al.

Developmental Cell, Journal Year: 2023, Volume and Issue: 58(23), P. 2641 - 2651.e6

Published: Oct. 26, 2023

Language: Английский

Citations

10
A Major Disease-Related Point Mutation in Spastin Dramatically Alters the Dynamics and Allostery of the Motor DOI
Shehani Kahawatte, Amanda C. Macke,

Carter St. Clair

et al.

Biochemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 26, 2025

Spastin is a microtubule-severing AAA+ ATPase that highly expressed in neuronal cells and plays crucial role axonal growth, branching, regeneration. This machine oligomerizes into hexamers the presence of ATP microtubule carboxy-terminal tails (CTTs). Conformational changes spastin hexamers, powered by hydrolysis, apply forces to microtubule, ultimately leading severing filament. Mutations disrupt normal function spastin, impairing its ability sever microtubules effectively abnormal dynamics neurons characteristic set neurodegenerative disorders called hereditary spastic paraplegias (HSP). Experimental studies have identified HSP-related R591S (Drosophila melanogaster numbering) mutation as playing spastin. Given significant HSP, we employed combination molecular simulations with learning graph network-based approaches identify quantify perturbations caused HSP on spastin's allostery functional implications. We found hexamer, upon mutation, loses execute primary motion associated action. The study allosteric showed regions are most perturbed those involved formation interprotomer contacts. induces rigidity networks motor, making it more likely experience loss applied would not be easily dissipated passing through variety alternative paths wild-type (WT)

Language: Английский

Citations

0
UCHL1-Mediated Spastin Degradation Regulates Microtubule Severing and Hippocampal Neurite Outgrowth DOI
Ao Ma, Zhi Liang, Hongde Zhang

et al.

Journal of Molecular Neuroscience, Journal Year: 2025, Volume and Issue: 75(2)

Published: April 24, 2025

Language: Английский

Citations

0
14-3-3 protein augments the protein stability of phosphorylated spastin and promotes the recovery of spinal cord injury through its agonist intervention DOI Creative Commons
Qiuling Liu, Hua Yang, Jianxian Luo

et al.

eLife, Journal Year: 2023, Volume and Issue: 12

Published: Sept. 12, 2023

Axon regeneration is abortive in the central nervous system following injury. Orchestrating microtubule dynamics has emerged as a promising approach to improve axonal regeneration. The severing enzyme spastin essential for development and through remodeling of arrangement. To date, however, little known regarding mechanisms underlying action neural after spinal cord Here, we use glutathione transferase pulldown immunoprecipitation assays demonstrate that 14-3-3 interacts with spastin, both vivo vitro, via Ser233 phosphorylation. Moreover, show protects from degradation by inhibiting ubiquitination pathway upregulates spastin-dependent ability. Furthermore, agonist Fusicoccin (FC-A) promotes neurite outgrowth vitro which needs activation. Western blot immunofluorescence results revealed protein upregulated neuronal compartment injury vivo. In addition, administration FC-A not only locomotor recovery, but also nerve contusion lateral hemisection models; application inhibitor spastazoline successfully reverses these phenomena. Taken together, indicate molecular switch regulates levels, small molecule effectively mediates recovery mice requires participation.

Language: Английский

Citations

5
Doublecortin restricts neuronal branching by regulating tubulin polyglutamylation DOI Creative Commons
Muriel Sébastien, Alexandra Paquette, Emily N. P. Prowse

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: June 3, 2023

Doublecortin (DCX) is a neuronal microtubule-associated protein (MAP) that binds directly to microtubules via two (DC) domains. The DC domains sense the nucleotide state, longitudinal curvature, and protofilament number of microtubule lattice, indicating role in regulation structure neurons. Mutations DCX cause lissencephaly subcortical band heterotopia (also known as double-cortex syndrome) due impaired migration. To better understand migration, we developed model system based on induced pluripotent stem cells (iPSCs). We used CRISPR/Cas9 knock-out Dcx gene iPSCs differentiated into cortical Compared control neurons, DCX-KO neurons showed reduced velocities nuclear movements. coincided with an increase neurites early development, consistent migration phenotype previous findings mouse model. Neurite branching regulated by host MAPs other factors, well polymerization dynamics. However, EB comet dynamics were unchanged similar growth rates, lifetimes, numbers. Rather, observed significant reduction α-tubulin polyglutamylation Polyglutamylation levels rescued expression or TTLL11, glutamylase. Using U2OS orthogonal system, show TTLL11 act synergistically promote polyglutamylation. regulates numerous MAPs, severing enzymes, molecular motors. Consistently, observe lysosomes their processivity. propose acts positive regulator restricts neurite branching. Our results indicate unexpected for homeostasis tubulin code.

Language: Английский

Citations

4
Unraveling Isoform Complexity: The Roles of M1‐ and M87‐Spastin in Spastic Paraplegia 4 (SPG4) DOI
S. Ramakrishnan, Neha Mohan, Zhangji Dong

et al.

Movement Disorders, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 29, 2024

Abstract Spastic Paraplegia 4 (SPG4) is a debilitating neurodegenerative disorder characterized by progressive muscle weakness and spasticity in the lower limbs, often leading to gait impairment. Central SPG4 pathology die‐back degeneration of corticospinal tracts, primarily driven mutations spastin protein encoded SPAST gene. gives rise two major isoforms, M1‐ M87‐spastin, which are generated from distinct translation initiation sites. Although implicated various cellular functions, specific roles each isoform pathogenesis remain poorly understood. This review offers an overview genetic structural organization M87‐spastin highlighting their overlapping exploring potential haploinsufficiency gain‐of‐toxicity mechanisms underlying SPG4. We also present novel perspective on evolutionary emergence M1‐spastin its unique involvement Drawing upon latest research, we propose intriguing hypothesis regarding hetero‐oligomerization how interaction may drive disease progression open new avenues for therapeutic intervention. By integrating current research with these fresh insights, seek illuminate complex molecular driving foster development innovative strategies. not only incorporates existing knowledge but lays groundwork future studies aimed at uncovering isoform‐specific SPG4, ultimate goal advancing targeted treatments this challenging disorder. © 2024 International Parkinson Movement Disorder Society.

Language: Английский

Citations

1
From spastic paraplegia to infantile neurodegenerative disorder: Expanding the phenotypic spectrum associated with biallelic SPAST variants DOI Creative Commons

Manon Degoutin,

Chloé Angelini, Claire Bar

et al.

European Journal of Neurology, Journal Year: 2024, Volume and Issue: 32(1)

Published: Dec. 27, 2024

Heterozygous pathogenic variants in SPAST are known to cause Hereditary Spastic Paraplegia 4 (SPG4), the most common form of HSP, characterized by progressive bilateral lower limbs spasticity with frequent sphincter disorders. However, there very few descriptions literature patients carrying biallelic SPAST. Targeted Sanger sequencing, panel sequencing and exome were used identify genetic causes 9 from 6 unrelated families symptoms HSP or infantile neurodegenerative disorder. We describe 5 pure a variable age onset, mostly infancy, profound intellectual disability progressively worsening tetrapyramidal syndrome. The patients' parents, heterozygous carriers variants, included both asymptomatic classic forms SPG4. Biallelic may explain cases hereditary spastic paraplegia autosomal recessive inheritance. Furthermore, some also psychomotor regression an disorder, associated syndrome, new phenotype gene.

Language: Английский

Citations

1