Show simple item record

dc.contributor.authorTousley, Adelaide
dc.contributor.authorIuliano, Maria
dc.contributor.authorWeisman, Elizabeth
dc.contributor.authorSapp, Ellen
dc.contributor.authorRichardson, Heather
dc.contributor.authorVodicka, Petr
dc.contributor.authorAlexander, Jonathan
dc.contributor.authorAronin, Neil
dc.contributor.authorDiFiglia, Marian
dc.contributor.authorKegel-Gleason, Kimberly B.
dc.date2022-08-11T08:09:52.000
dc.date.accessioned2022-08-23T16:46:37Z
dc.date.available2022-08-23T16:46:37Z
dc.date.issued2019-02-15
dc.date.submitted2019-02-28
dc.identifier.citation<p>PLoS One. 2019 Feb 15;14(2):e0212337. doi: 10.1371/journal.pone.0212337. eCollection 2019. <a href="https://doi.org/10.1371/journal.pone.0212337">Link to article on publisher's site</a></p>
dc.identifier.issn1932-6203 (Linking)
dc.identifier.doi10.1371/journal.pone.0212337
dc.identifier.pmid30768638
dc.identifier.urihttp://hdl.handle.net/20.500.14038/40917
dc.description.abstractOne 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.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=30768638&dopt=Abstract">Link to Article in PubMed</a></p>
dc.rightsCopyright: © 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.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectFibroblasts
dc.subjectSmall interfering RNAs
dc.subjectCell membranes
dc.subjectActins
dc.subjectCell staining
dc.subjectFocal adhesions
dc.subjectImmunoprecipitation
dc.subjectImmunofluorescence
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectCell Biology
dc.subjectCells
dc.subjectGenetic Phenomena
dc.subjectNervous System Diseases
dc.subjectNeuroscience and Neurobiology
dc.subjectNucleic Acids, Nucleotides, and Nucleosides
dc.titleHuntingtin associates with the actin cytoskeleton and alpha-actinin isoforms to influence stimulus dependent morphology changes
dc.typeJournal Article
dc.source.journaltitlePloS one
dc.source.volume14
dc.source.issue2
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=4729&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/3717
dc.identifier.contextkey13921322
refterms.dateFOA2022-08-23T16:46:37Z
html.description.abstract<p>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.</p>
dc.identifier.submissionpathoapubs/3717
dc.contributor.departmentDepartment of Medicine, Division of Endocrinology and Metabolism
dc.source.pagese0212337


Files in this item

Thumbnail
Name:
journal.pone.0212337.pdf
Size:
4.550Mb
Format:
PDF

This item appears in the following Collection(s)

Show simple item record

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.
Except where otherwise noted, this item's license is described as 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.