The Plasma Membrane-Associated GTPase Rin Interacts with the Dopamine Transporter and Is Required for Protein Kinase C-Regulated Dopamine Transporter Trafficking
Authors
Navaroli, Deanna M.Stevens, Zachary H.
Uzelac, Zeljko
Gabriel, Luke
King, Michael J.
Lifshitz, Lawrence M.
Sitte, Harald H.
Melikian, Haley E.
Student Authors
Deanna M. Navaroli; Luke GabrielAcademic Program
NeuroscienceUMass Chan Affiliations
Melikian LabGraduate School of Biomedical Sciences, Neuroscience Program
Program in Molecular Medicine
Department of Psychiatry
Document Type
Journal ArticlePublication Date
2011-09-28Keywords
Dopamine Plasma Membrane Transport Proteins; ras ProteinsUMCCTS funding
Life Sciences
Medicine and Health Sciences
Neuroscience and Neurobiology
Metadata
Show full item recordAbstract
Dopaminergic signaling and plasticity are essential to numerous CNS functions and pathologies, including movement, cognition, and addiction. The amphetamine- and cocaine-sensitive dopamine (DA) transporter (DAT) tightly controls extracellular DA concentrations and half-life. DAT function and surface expression are not static but are dynamically modulated by membrane trafficking. We recently demonstrated that the DAT C terminus encodes a PKC-sensitive internalization signal that also suppresses basal DAT endocytosis. However, the cellular machinery governing regulated DAT trafficking is not well defined. In work presented here, we identified the Ras-like GTPase, Rin (for Ras-like in neurons) (Rit2), as a protein that interacts with the DAT C-terminal endocytic signal. Yeast two-hybrid, GST pull down and FRET studies establish that DAT and Rin directly interact, and colocalization studies reveal that DAT/Rin associations occur primarily in lipid raft microdomains. Coimmunoprecipitations demonstrate that PKC activation regulates Rin association with DAT. Perturbation of Rin function with GTPase mutants and shRNA-mediated Rin knockdown reveals that Rin is critical for PKC-mediated DAT internalization and functional downregulation. These results establish that Rin is a DAT-interacting protein that is required for PKC-regulated DAT trafficking. Moreover, this work suggests that Rin participates in regulated endocytosis.Source
J Neurosci. 2011 Sep 28;31(39):13758-70. Link to article on publisher's website
DOI
10.1523/JNEUROSCI.2649-11.2011Permanent Link to this Item
http://hdl.handle.net/20.500.14038/33221PubMed ID
21957239Related Resources
Link to article in PubMedae974a485f413a2113503eed53cd6c53
10.1523/JNEUROSCI.2649-11.2011