Department of Medicine, Division of Endocrinology and Metabolism
Amino Acids, Peptides, and Proteins | Cell Biology | Cells | Genetic Phenomena | Nervous System Diseases | Neuroscience and Neurobiology | Nucleic Acids, Nucleotides, and Nucleosides
One response of cells to growth factor stimulus involves changes in morphology driven by the actin cytoskeleton and actin associated proteins which regulate functions such as cell adhesion, motility and in neurons, synaptic plasticity. Previous studies suggest that Huntingtin may be involved in regulating morphology however, there has been limited evidence linking endogenous Huntingtin localization or function with cytoplasmic actin in cells. We found that depletion of Huntingtin in human fibroblasts reduced adhesion and altered morphology and these phenotypes were made worse with growth factor stimulation, whereas the presence of the Huntington's Disease mutation inhibited growth factor induced changes in morphology and increased numbers of vinculin-positive focal adhesions. Huntingtin immunoreactivity localized to actin stress fibers, vinculin-positive adhesion contacts and membrane ruffles in fibroblasts. Interactome data from others has shown that Huntingtin can associate with alpha-actinin isoforms which bind actin filaments. Mapping studies using a cDNA encoding alpha-actinin-2 showed that it interacts within Huntingtin aa 399-969. Double-label immunofluorescence showed Huntingtin and alpha-actinin-1 co-localized to stress fibers, membrane ruffles and lamellar protrusions in fibroblasts. Proximity ligation assays confirmed a close molecular interaction between Huntingtin and alpha-actinin-1 in human fibroblasts and neurons. Huntingtin silencing with siRNA in fibroblasts blocked the recruitment of alpha-actinin-1 to membrane foci. These studies support the idea that Huntingtin is involved in regulating adhesion and actin dependent functions including those involving alpha-actinin.
Fibroblasts, Small interfering RNAs, Cell membranes, Actins, Cell staining, Focal adhesions, Immunoprecipitation, Immunofluorescence
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Copyright: © 2019 Tousley et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
DOI of Published Version
PLoS One. 2019 Feb 15;14(2):e0212337. doi: 10.1371/journal.pone.0212337. eCollection 2019. Link to article on publisher's site
Tousley, Adelaide; Iuliano, Maria; Weisman, Elizabeth; Sapp, Ellen; Richardson, Heather; Vodicka, Petr; Alexander, Jonathan; Aronin, Neil; DiFiglia, Marian; and Kegel-Gleason, Kimberly B., "Huntingtin associates with the actin cytoskeleton and alpha-actinin isoforms to influence stimulus dependent morphology changes" (2019). Open Access Articles. 3717.
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