Phosphorylation of homer3 by calcium/calmodulin-dependent kinase II regulates a coupling state of its target molecules in purkinje cells

Akihiro Mizutani, Yukiko Kuroda, Akira Futatsugi, Teiichi Furuichi, Katsuhiko Mikoshiba

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Homer proteins are components of postsynaptic density (PSD) and play a crucial role in coupling diverse target molecules. However, the regulatory aspect of Homer-mediated coupling has been addressed only about a dominant-negative effect of Homer1a, which requires de novo gene expression. Here, we present evidence that Homer-mediated coupling is regulated by its phosphorylation state. We found that Homer3, the predominant isoform in Purkinje cells, is phosphorylated by calcium/calmodulin-dependent protein kinase II (CaMKII) both in vitro and in vivo. Biochemical fractionation with phosphor-specific antibodies revealed the presence of phosphorylated Homer3 in the cytosolic fraction in contrast to high levels of nonphosphorylated Homer3 in PSD. In P/Q-type voltage-gated-Ca2+ channel knock-out mice, in which CaMKII activation was reduced, the levels of Homer3 phosphorylation and the soluble form of Homer 3 were markedly lower. Furthermore, both robust phosphorylation of Homer3 and its dissociation from metabotropic glutamate receptor 1α (mGluR1α) were triggered by depolarization in primary cultured Purkinje cells, and these events were inhibited by CaMKII inhibitor. An in vitro binding kinetic analysis revealed that these phosphorylation-dependent events were attributable to a decrease in the affinity of phosphorylated Homer3 for its ligand. In a heterologous system, the Ca2+ signaling pattern induced by mGluR1α activation was modulated by the Homer3 phosphorylation state. Together, these findings suggested that Homer3 in Purkinje cells might function as a reversible coupler regulated by CaMKII phosphorylation and that the phosphorylation is capable of regulating the postsynaptic molecular architecture in response to synaptic activity.

Original languageEnglish
Pages (from-to)5369-5382
Number of pages14
JournalJournal of Neuroscience
Volume28
Issue number20
DOIs
Publication statusPublished - 2008 May 14
Externally publishedYes

Fingerprint

Calcium-Calmodulin-Dependent Protein Kinases
Purkinje Cells
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Phosphorylation
Calcium
Post-Synaptic Density
Protein Kinase Inhibitors
Knockout Mice
Cultured Cells
Protein Isoforms
Ligands
Gene Expression
Antibodies

Keywords

  • Ca signaling
  • CaMKII
  • Homer
  • Phosphorylation
  • Purkinje cells
  • Synaptic plasticity

ASJC Scopus subject areas

  • Neuroscience(all)
  • Medicine(all)

Cite this

Phosphorylation of homer3 by calcium/calmodulin-dependent kinase II regulates a coupling state of its target molecules in purkinje cells. / Mizutani, Akihiro; Kuroda, Yukiko; Futatsugi, Akira; Furuichi, Teiichi; Mikoshiba, Katsuhiko.

In: Journal of Neuroscience, Vol. 28, No. 20, 14.05.2008, p. 5369-5382.

Research output: Contribution to journalArticle

Mizutani, Akihiro ; Kuroda, Yukiko ; Futatsugi, Akira ; Furuichi, Teiichi ; Mikoshiba, Katsuhiko. / Phosphorylation of homer3 by calcium/calmodulin-dependent kinase II regulates a coupling state of its target molecules in purkinje cells. In: Journal of Neuroscience. 2008 ; Vol. 28, No. 20. pp. 5369-5382.
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