Inhibition of Dopamine Transporter Activity by G Protein βγ Subunits

Jennie Garcia-Olivares, Delany Torres-Salazar, William A. Owens, Tracy Baust, David P. Siderovski, Susan G. Amara, Jun Zhu, Lynette C Daws, Gonzalo E. Torres

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Uptake through the Dopamine Transporter (DAT) is the primary mechanism of terminating dopamine signaling within the brain, thus playing an essential role in neuronal homeostasis. Deregulation of DAT function has been linked to several neurological and psychiatric disorders including ADHD, schizophrenia, Parkinson's disease, and drug addiction. Over the last 15 years, several studies have revealed a plethora of mechanisms influencing the activity and cellular distribution of DAT; suggesting that fine-tuning of dopamine homeostasis occurs via an elaborate interplay of multiple pathways. Here, we show for the first time that the βγ subunits of G proteins regulate DAT activity. In heterologous cells and brain tissue, a physical association between Gβγ subunits and DAT was demonstrated by co-immunoprecipitation. Furthermore, in vitro pull-down assays using purified proteins established that this association occurs via a direct interaction between the intracellular carboxy-terminus of DAT and Gβγ. Functional assays performed in the presence of the non-hydrolyzable GTP analog GTP-γ-S, Gβγ subunit overexpression, or the Gβγ activator mSIRK all resulted in rapid inhibition of DAT activity in heterologous systems. Gβγ activation by mSIRK also inhibited dopamine uptake in brain synaptosomes and dopamine clearance from mouse striatum as measured by high-speed chronoamperometry in vivo. Gβγ subunits are intracellular signaling molecules that regulate a multitude of physiological processes through interactions with enzymes and ion channels. Our findings add neurotransmitter transporters to the growing list of molecules regulated by G-proteins and suggest a novel role for Gβγ signaling in the control of dopamine homeostasis.

Original languageEnglish (US)
Article numbere59788
JournalPLoS One
Volume8
Issue number3
DOIs
StatePublished - Mar 26 2013

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Dopamine Plasma Membrane Transport Proteins
Protein Subunits
protein subunits
dopamine
G-proteins
GTP-Binding Proteins
transporters
Dopamine
Brain
Homeostasis
Guanosine Triphosphate
Assays
Neurotransmitter Transport Proteins
homeostasis
Association reactions
Physiological Phenomena
Chronoamperometry
Molecules
Deregulation
Synaptosomes

ASJC Scopus subject areas

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

Cite this

Garcia-Olivares, J., Torres-Salazar, D., Owens, W. A., Baust, T., Siderovski, D. P., Amara, S. G., ... Torres, G. E. (2013). Inhibition of Dopamine Transporter Activity by G Protein βγ Subunits. PLoS One, 8(3), [e59788]. https://doi.org/10.1371/journal.pone.0059788

Inhibition of Dopamine Transporter Activity by G Protein βγ Subunits. / Garcia-Olivares, Jennie; Torres-Salazar, Delany; Owens, William A.; Baust, Tracy; Siderovski, David P.; Amara, Susan G.; Zhu, Jun; Daws, Lynette C; Torres, Gonzalo E.

In: PLoS One, Vol. 8, No. 3, e59788, 26.03.2013.

Research output: Contribution to journalArticle

Garcia-Olivares, J, Torres-Salazar, D, Owens, WA, Baust, T, Siderovski, DP, Amara, SG, Zhu, J, Daws, LC & Torres, GE 2013, 'Inhibition of Dopamine Transporter Activity by G Protein βγ Subunits', PLoS One, vol. 8, no. 3, e59788. https://doi.org/10.1371/journal.pone.0059788
Garcia-Olivares J, Torres-Salazar D, Owens WA, Baust T, Siderovski DP, Amara SG et al. Inhibition of Dopamine Transporter Activity by G Protein βγ Subunits. PLoS One. 2013 Mar 26;8(3). e59788. https://doi.org/10.1371/journal.pone.0059788
Garcia-Olivares, Jennie ; Torres-Salazar, Delany ; Owens, William A. ; Baust, Tracy ; Siderovski, David P. ; Amara, Susan G. ; Zhu, Jun ; Daws, Lynette C ; Torres, Gonzalo E. / Inhibition of Dopamine Transporter Activity by G Protein βγ Subunits. In: PLoS One. 2013 ; Vol. 8, No. 3.
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