DHHC5 protein palmitoylates flotillin-2 and is rapidly degraded on induction of neuronal differentiation in cultured cells

Yi Li, Brent R. Martin, Benjamin F. Cravatt, Sandra L. Hofmann

Research output: Contribution to journalArticle

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Abstract

Post-translational palmitoylation of intracellular proteins is mediated by protein palmitoyltransferases belonging to the DHHC family, which share a common catalytic Asp-His-His-Cys (DHHC) motif. Several members have been implicated in neuronal development, neurotransmission, and synaptic plasticity. We previously observed that mice homozygous for a hypomorphic allele of the ZDHHC5 gene are impaired in context-dependent learning and memory. To identify potentially relevant protein substrates of DHHC5, we performed a quantitative proteomic analysis of stable isotope-labeled neuronal stem cell cultures from forebrains of normal andDHHC5-GT(gene-trapped) mice using the bioorthogonal palmitate analog 17-octadecynoic acid.We identified ∼300 17-octadecynoic acid-modified and hydroxylamine-sensitive proteins, of which a subset was decreased in abundance inDHHC5-GTcells. Palmitoylation and oligomerization of one of these proteins (flotillin- 2) was abolished in DHHC5-GT neuronal stem cells. In COS-1 cells, overexpression of DHHC5 markedly stimulated the palmitoylation of flotillin-2, strongly suggesting a direct enzyme-substrate relationship. Serendipitously, we found that down-regulation of DHHC5 was triggered within minutes following growth factor withdrawal from normal neural stem cells, a maneuver that is used to induce neural differentiation in culture. The effect was reversible for up to 4 h, and degradation was partially prevented by inhibitors of ubiquitin-mediated proteolysis. These findings suggest that protein palmitoylation can be regulated through changes in DHHC PAT levels in response to differentiation signals.

Original languageEnglish (US)
Pages (from-to)523-530
Number of pages8
JournalJournal of Biological Chemistry
Volume287
Issue number1
DOIs
StatePublished - Jan 2 2012

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Lipoylation
Cultured Cells
Cells
Stem cells
Proteins
Stem Cells
Genes
Proteolysis
Oligomerization
Hydroxylamine
Neuronal Plasticity
Neural Stem Cells
Palmitates
COS Cells
Corrosion inhibitors
Substrates
Prosencephalon
Ubiquitin
Cell culture
Synaptic Transmission

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

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DHHC5 protein palmitoylates flotillin-2 and is rapidly degraded on induction of neuronal differentiation in cultured cells. / Li, Yi; Martin, Brent R.; Cravatt, Benjamin F.; Hofmann, Sandra L.

In: Journal of Biological Chemistry, Vol. 287, No. 1, 02.01.2012, p. 523-530.

Research output: Contribution to journalArticle

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