Phosphorylation of mutant huntingtin at serine 116 modulates neuronal toxicity

Erin E. Watkin, Nicolas Arbez, Elaine Waldron-Roby, Robert O'Meally, Tamara Ratovitski, Robert N Cole, Christopher A Ross

Research output: Contribution to journalArticle

Abstract

Phosphorylation has been shown to have a significant impact on expanded huntingtin-mediated cellular toxicity. Several phosphorylation sites have been identified on the huntingtin (Htt) protein. To find new potential therapeutic targets for Huntington's Disease (HD), we used mass spectrometry to identify novel phosphorylation sites on N-terminal Htt, expressed in HEK293 cells. Using site-directed mutagenesis we introduced alterations of phosphorylation sites in a N586 Htt construct containing 82 polyglutamine repeats. The effects of these alterations on expanded Htt toxicity were evaluated in primary neurons using a nuclear condensation assay and a direct time-lapse imaging of neuronal death. As a result of these studies, we identified several novel phosphorylation sites, validated several known sites, and discovered one phospho-null alteration, S116A, that had a protective effect against expanded polyglutamine-mediated cellular toxicity. The results suggest that S116 is a potential therapeutic target, and indicate that our screening method is useful for identifying candidate phosphorylation sites. Copyright:

Original languageEnglish (US)
Article numbere88284
JournalPLoS One
Volume9
Issue number2
DOIs
StatePublished - Feb 5 2014

Fingerprint

Phosphorylation
serine
Serine
Toxicity
phosphorylation
toxicity
mutants
cytotoxicity
Time-Lapse Imaging
therapeutics
Mutagenesis
HEK293 Cells
Huntington Disease
site-directed mutagenesis
Site-Directed Mutagenesis
Neurons
Mass spectrometry
protective effect
Condensation
Assays

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Phosphorylation of mutant huntingtin at serine 116 modulates neuronal toxicity. / Watkin, Erin E.; Arbez, Nicolas; Waldron-Roby, Elaine; O'Meally, Robert; Ratovitski, Tamara; Cole, Robert N; Ross, Christopher A.

In: PLoS One, Vol. 9, No. 2, e88284, 05.02.2014.

Research output: Contribution to journalArticle

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