RNA Therapeutics Institute; Department of Medicine
Amino Acids, Peptides, and Proteins | Biochemistry, Biophysics, and Structural Biology | Enzymes and Coenzymes | Nervous System Diseases | Neuroscience and Neurobiology | Nucleic Acids, Nucleotides, and Nucleosides | Physiology
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, p85alpha subunit of PI 3-kinase, and the actin bundling protein alpha-actinin and interacts preferentially with the GTP bound form of Rac1. The HD mutation reduced Huntingtin interaction with p85alpha.
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.
Actin, GTPase, HTT, PI 3-kinase, growth factor, signaling, “Ras-Related C3 Botulinum Toxin Substrate 1”
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Copyright © 2019 – IOS Press and the authors. This is an open access article distributed under the terms of the Creative Commons Attribution Non-Commercial (CC BY-NC 4.0) License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
DOI of Published Version
J Huntingtons Dis. 2019;8(1):53-69. doi: 10.3233/JHD-180311. Link to article on publisher's site
Journal of Huntington's disease
Tousley A, Khvorova A, Aronin N, Kegel-Gleason KB. (2019). Rac1 Activity Is Modulated by Huntingtin and Dysregulated in Models of Huntington's Disease. RNA Therapeutics Institute Publications. https://doi.org/10.3233/JHD-180311. Retrieved from https://escholarship.umassmed.edu/rti_pubs/48
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