The multiple LIM domain-containing adaptor protein Hic-5 synaptically colocalizes and interacts with the dopamine transporter

Ana M. Carneiro, Susan L. Ingram, Jean Martin Beaulieu, Ava Sweeney, Susan G. Amara, Sheila M. Thomas, Marc G. Caron, Gonzalo E. Torres

Research output: Contribution to journalArticle

90 Scopus citations

Abstract

The Na+/Cl--dependent dopamine transporter (DAT) is critical in terminating dopaminergic transmission by removing the transmitter away from the synapse. Several lines of evidence suggest that transporter-interacting proteins may play a role in DAT function and regulation. In this report, using the yeast two-hybrid system, we have identified a novel interaction between DAT and the multiple Lin-11, IsI-1, and Mec-3 (LIM) domain-containing adaptor protein Hic-5. This association involves the N-terminal portion of the intracellular tail of DAT and the LIM region of Hic-5. In human embryonic kidney 293 cells, Hic-5 colocalizes with DAT at polarized sites and reduces DAT uptake activity through a mechanism involving a decrease in the cell-surface levels of the transporter. A fragment of Hic-5 containing the LIM domains is sufficient to bind DAT but lacks the ability to inhibit transporter activity. In addition, the LIM fragment prevents the effect of the full-length Hic-5 on DAT localization and function. In the brain, Hic-5 protein is expressed in the cerebral cortex, hippocampus, hypothalamus, cerebellum, and striatum, suggesting a role for this protein in the nervous system. The association of the endogenous Hic-5 and DAT proteins was confirmed biochemically by coimmunoprecipitation from brain striatal extracts. Moreover, immunostaining of rat midbrain neurons in culture revealed a presynaptic colocalization of Hic-5 and DAT. Because Hic-5 has been shown to interact with several signaling molecules, including the nonreceptor protein tyrosine kinases focal adhesion kinase and Fyn, this raises the possibility that this adaptor protein may link DAT to intracellular signaling pathways.

Original languageEnglish (US)
Pages (from-to)7045-7054
Number of pages10
JournalJournal of Neuroscience
Volume22
Issue number16
StatePublished - Aug 15 2002

Keywords

  • Adaptor
  • Colocalization
  • Dopamine transporter
  • Hic-5
  • Interaction
  • LIM domain
  • Uptake

ASJC Scopus subject areas

  • Neuroscience(all)

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    Carneiro, A. M., Ingram, S. L., Beaulieu, J. M., Sweeney, A., Amara, S. G., Thomas, S. M., Caron, M. G., & Torres, G. E. (2002). The multiple LIM domain-containing adaptor protein Hic-5 synaptically colocalizes and interacts with the dopamine transporter. Journal of Neuroscience, 22(16), 7045-7054.