Rac1 Activity Is Modulated by Huntingtin and Dysregulated in Models of Huntington's Disease

J Huntingtons Dis. 2019;8(1):53-69. doi: 10.3233/JHD-180311.

Abstract

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.

Keywords: Actin; GTPase; HTT; PI 3-kinase; growth factor; signaling; “Ras-Related C3 Botulinum Toxin Substrate 1”.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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*

Substances

  • HTT protein, human
  • Huntingtin Protein
  • Microfilament Proteins
  • Nerve Tissue Proteins
  • Neuropeptides
  • RAC1 protein, human
  • Rac1 protein, mouse
  • rac1 GTP-Binding Protein