Dopamine Transporter Membrane Mobility is Bidirectionally Regulated by Phosphorylation and Palmitoylation

Overview

Journal Curr Res Physiol

Date 2023 Dec 18

PMID 38107792

Authors

Madhur Shetty

Danielle E Bolland

Joshua Morrell

Bryon D Grove

James D Foster

Roxanne A Vaughan

Affiliations

Soon will be listed here.

Abstract

The primary regulator of dopamine availability in the brain is the dopamine transporter (DAT), a plasma membrane protein that drives reuptake of released dopamine from the extracellular space into the presynaptic neuron. DAT activity is regulated by post-translational modifications that establish clearance capacity through impacts on transport kinetics, and dysregulation of these events may underlie dopaminergic imbalances in mood and psychiatric disorders. Here, using fluorescence recovery after photobleaching, we show that phosphorylation and palmitoylation induce opposing effects on DAT lateral membrane mobility, which may influence functional outcomes by regulating subcellular localization and binding partner interactions. Membrane mobility was also impacted by amphetamine and in polymorphic variant A559V in directions consistent with enhanced phosphorylation. These findings grow the list of DAT properties controlled by these post-translational modifications and highlight their role in establishment of dopaminergic tone in physiological and pathophysiological states.

Citing Articles

Recapitulating the potential contribution of protein S-palmitoylation in cancer.

Chaturvedi S, Sonawane A Cancer Metastasis Rev. 2024; 44(1):20.

PMID: 39725785 DOI: 10.1007/s10555-024-10217-3.

Altered Protein Palmitoylation as Disease Mechanism in Neurodegenerative Disorders.

Wlodarczyk J, Bhattacharyya R, Dore K, Ho G, Martin D, Mejias R J Neurosci. 2024; 44(40).

PMID: 39358031 PMC: 11450541. DOI: 10.1523/JNEUROSCI.1225-24.2024.

Fluorescence photobleaching and recovery of fluorescein sodium in carbomer film.

Lin Y, Chen H, Yang Y RSC Adv. 2024; 14(6):3841-3844.

PMID: 38274174 PMC: 10810102. DOI: 10.1039/d3ra08718b.

References

Miranda M, Wu C, Sorkina T, Korstjens D, Sorkin A . Enhanced ubiquitylation and accelerated degradation of the dopamine transporter mediated by protein kinase C. J Biol Chem. 2005; 280(42):35617-24. DOI: 10.1074/jbc.M506618200. View

Rahbek-Clemmensen T, Lycas M, Erlendsson S, Eriksen J, Apuschkin M, Vilhardt F . Super-resolution microscopy reveals functional organization of dopamine transporters into cholesterol and neuronal activity-dependent nanodomains. Nat Commun. 2017; 8(1):740. PMC: 5622129. DOI: 10.1038/s41467-017-00790-3. View

Fukata M, Fukata Y, Adesnik H, Nicoll R, Bredt D . Identification of PSD-95 palmitoylating enzymes. Neuron. 2004; 44(6):987-96. DOI: 10.1016/j.neuron.2004.12.005. View

Cervinski M, Foster J, Vaughan R . Psychoactive substrates stimulate dopamine transporter phosphorylation and down-regulation by cocaine-sensitive and protein kinase C-dependent mechanisms. J Biol Chem. 2005; 280(49):40442-9. DOI: 10.1074/jbc.M501969200. View

Kang M, Day C, Kenworthy A, DiBenedetto E . Simplified equation to extract diffusion coefficients from confocal FRAP data. Traffic. 2012; 13(12):1589-600. PMC: 3731631. DOI: 10.1111/tra.12008. View

Cremona M, Matthies H, Pau K, Bowton E, Speed N, Lute B . Flotillin-1 is essential for PKC-triggered endocytosis and membrane microdomain localization of DAT. Nat Neurosci. 2011; 14(4):469-77. PMC: 3066276. DOI: 10.1038/nn.2781. View

Thal L, Tomlinson I, Quinlan M, Kovtun O, Blakely R, Rosenthal S . Single Quantum Dot Imaging Reveals PKCβ-Dependent Alterations in Membrane Diffusion and Clustering of an Attention-Deficit Hyperactivity Disorder/Autism/Bipolar Disorder-Associated Dopamine Transporter Variant. ACS Chem Neurosci. 2018; 10(1):460-471. PMC: 6411462. DOI: 10.1021/acschemneuro.8b00350. View

Piniella D, Martinez-Blanco E, Bartolome-Martin D, Sanz-Martos A, Zafra F . Identification by proximity labeling of novel lipidic and proteinaceous potential partners of the dopamine transporter. Cell Mol Life Sci. 2021; 78(23):7733-7756. PMC: 8629785. DOI: 10.1007/s00018-021-03998-1. View

Thompson A, Zapata A, Justice Jr J, Vaughan R, SHARPE L, Shippenberg T . Kappa-opioid receptor activation modifies dopamine uptake in the nucleus accumbens and opposes the effects of cocaine. J Neurosci. 2000; 20(24):9333-40. PMC: 6773021. View

10.

Bowton E, Saunders C, Reddy I, Campbell N, Hamilton P, Henry L . SLC6A3 coding variant Ala559Val found in two autism probands alters dopamine transporter function and trafficking. Transl Psychiatry. 2014; 4:e464. PMC: 4350523. DOI: 10.1038/tp.2014.90. View

11.

Foster J, Vaughan R . Palmitoylation controls dopamine transporter kinetics, degradation, and protein kinase C-dependent regulation. J Biol Chem. 2010; 286(7):5175-86. PMC: 3037630. DOI: 10.1074/jbc.M110.187872. View

12.

Lee K, Kim M, Kim D, Lee Y . Syntaxin 1A and receptor for activated C kinase interact with the N-terminal region of human dopamine transporter. Neurochem Res. 2004; 29(7):1405-9. DOI: 10.1023/b:nere.0000026404.08779.43. View

13.

Huff R, Vaughan R, Kuhar M, Uhl G . Phorbol esters increase dopamine transporter phosphorylation and decrease transport Vmax. J Neurochem. 1997; 68(1):225-32. DOI: 10.1046/j.1471-4159.1997.68010225.x. View

14.

Wetzker R, Bohmer F . Transactivation joins multiple tracks to the ERK/MAPK cascade. Nat Rev Mol Cell Biol. 2003; 4(8):651-7. DOI: 10.1038/nrm1173. View

15.

Vaughan R, Foster J . Mechanisms of dopamine transporter regulation in normal and disease states. Trends Pharmacol Sci. 2013; 34(9):489-96. PMC: 3831354. DOI: 10.1016/j.tips.2013.07.005. View

16.

Foster J, Pananusorn B, Vaughan R . Dopamine transporters are phosphorylated on N-terminal serines in rat striatum. J Biol Chem. 2002; 277(28):25178-86. DOI: 10.1074/jbc.M200294200. View

17.

Bolland D, Moritz A, Stanislowski D, Vaughan R, Foster J . Palmitoylation by Multiple DHHC Enzymes Enhances Dopamine Transporter Function and Stability. ACS Chem Neurosci. 2019; 10(6):2707-2717. PMC: 6746250. DOI: 10.1021/acschemneuro.8b00558. View

18.

Foster J, Adkins S, Lever J, Vaughan R . Phorbol ester induced trafficking-independent regulation and enhanced phosphorylation of the dopamine transporter associated with membrane rafts and cholesterol. J Neurochem. 2008; 105(5):1683-99. PMC: 8981492. DOI: 10.1111/j.1471-4159.2008.05262.x. View

19.

Torres G, Yao W, Mohn A, Quan H, Kim K, Levey A . Functional interaction between monoamine plasma membrane transporters and the synaptic PDZ domain-containing protein PICK1. Neuron. 2001; 30(1):121-34. DOI: 10.1016/s0896-6273(01)00267-7. View

20.

Eriksen J, Rasmussen S, Rasmussen T, Vaegter C, Cha J, Zou M . Visualization of dopamine transporter trafficking in live neurons by use of fluorescent cocaine analogs. J Neurosci. 2009; 29(21):6794-808. PMC: 3849467. DOI: 10.1523/JNEUROSCI.4177-08.2009. View