Rac1 Activity Is Modulated by Huntingtin and Dysregulated in Models of Huntington's Disease.
Animals
Cell Differentiation
Corpus Striatum
/ metabolism
Disease Models, Animal
Humans
Huntingtin Protein
/ genetics
Huntington Disease
/ genetics
Mice
Microfilament Proteins
/ metabolism
Mutation
/ genetics
Nerve Tissue Proteins
/ genetics
Neurons
/ metabolism
Neuropeptides
/ genetics
Signal Transduction
/ genetics
rac1 GTP-Binding Protein
/ genetics
Actin
GTPase
HTT
PI 3-kinase
growth factor
signaling
“Ras-Related C3 Botulinum Toxin Substrate 1”
Journal
Journal of Huntington's disease
ISSN: 1879-6400
Titre abrégé: J Huntingtons Dis
Pays: Netherlands
ID NLM: 101589965
Informations de publication
Date de publication:
2019
2019
Historique:
pubmed:
31
12
2018
medline:
13
3
2020
entrez:
31
12
2018
Statut:
ppublish
Résumé
Previous studies suggest that Huntingtin, the protein mutated in Huntington's disease (HD), is required for actin based changes in cell morphology, and undergoes stimulus induced targeting to plasma membranes where it interacts with phospholipids involved in cell signaling. The small GTPase Rac1 is a downstream target of growth factor stimulation and PI 3-kinase activity and is critical for actin dependent membrane remodeling. To determine if Rac1 activity is impaired in HD or regulated by normal Huntingtin. Analyses were performed in differentiated control and HD human stem cells and HD Q140/Q140 knock-in mice. Biochemical methods included SDS-PAGE, western blot, immunoprecipitation, affinity chromatography, and ELISA based Rac activity assays. Basal Rac1 activity increased following depletion of Huntingtin with Huntingtin specific siRNA in human primary fibroblasts and in human control neuron cultures. Human cells (fibroblasts, neural stem cells, and neurons) with the HD mutation failed to increase Rac1 activity in response to growth factors. Rac1 activity levels were elevated in striatum of 1.5-month-old HD Q140/Q140 mice and in primary embryonic cortical neurons from HD mice. Affinity chromatography analysis of striatal lysates showed that Huntingtin is in a complex with Rac1, p85α subunit of PI 3-kinase, and the actin bundling protein α-actinin and interacts preferentially with the GTP bound form of Rac1. The HD mutation reduced Huntingtin interaction with p85α. These findings suggest that Huntingtin regulates Rac1 activity as part of a coordinated response to growth factor signaling and this function is impaired early in HD.
Sections du résumé
BACKGROUND
Previous studies suggest that Huntingtin, the protein mutated in Huntington's disease (HD), is required for actin based changes in cell morphology, and undergoes stimulus induced targeting to plasma membranes where it interacts with phospholipids involved in cell signaling. The small GTPase Rac1 is a downstream target of growth factor stimulation and PI 3-kinase activity and is critical for actin dependent membrane remodeling.
OBJECTIVE
To determine if Rac1 activity is impaired in HD or regulated by normal Huntingtin.
METHODS
Analyses were performed in differentiated control and HD human stem cells and HD Q140/Q140 knock-in mice. Biochemical methods included SDS-PAGE, western blot, immunoprecipitation, affinity chromatography, and ELISA based Rac activity assays.
RESULTS
Basal Rac1 activity increased following depletion of Huntingtin with Huntingtin specific siRNA in human primary fibroblasts and in human control neuron cultures. Human cells (fibroblasts, neural stem cells, and neurons) with the HD mutation failed to increase Rac1 activity in response to growth factors. Rac1 activity levels were elevated in striatum of 1.5-month-old HD Q140/Q140 mice and in primary embryonic cortical neurons from HD mice. Affinity chromatography analysis of striatal lysates showed that Huntingtin is in a complex with Rac1, p85α subunit of PI 3-kinase, and the actin bundling protein α-actinin and interacts preferentially with the GTP bound form of Rac1. The HD mutation reduced Huntingtin interaction with p85α.
CONCLUSIONS
These findings suggest that Huntingtin regulates Rac1 activity as part of a coordinated response to growth factor signaling and this function is impaired early in HD.
Identifiants
pubmed: 30594931
pii: JHD180311
doi: 10.3233/JHD-180311
pmc: PMC6398565
doi:
Substances chimiques
HTT protein, human
0
Huntingtin Protein
0
Microfilament Proteins
0
Nerve Tissue Proteins
0
Neuropeptides
0
RAC1 protein, human
0
Rac1 protein, mouse
0
rac1 GTP-Binding Protein
EC 3.6.5.2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
53-69Subventions
Organisme : NINDS NIH HHS
ID : R01 NS100529
Pays : United States
Organisme : NIH HHS
ID : S10 OD020012
Pays : United States
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