---
_id: '7880'
abstract:
- lang: eng
  text: 'Following its evoked release, dopamine (DA) signaling is rapidly terminated
    by presynaptic reuptake, mediated by the cocaine-sensitive DA transporter (DAT).
    DAT surface availability is dynamically regulated by endocytic trafficking, and
    direct protein kinase C (PKC) activation acutely diminishes DAT surface expression
    by accelerating DAT internalization. Previous cell line studies demonstrated that
    PKC-stimulated DAT endocytosis requires both Ack1 inactivation, which releases
    a DAT-specific endocytic brake, and the neuronal GTPase, Rit2, which binds DAT.
    However, it is unknown whether Rit2 is required for PKC-stimulated DAT endocytosis
    in DAergic terminals or whether there are region- and/or sex-dependent differences
    in PKC-stimulated DAT trafficking. Moreover, the mechanisms by which Rit2 controls
    PKC-stimulated DAT endocytosis are unknown. Here, we directly examined these important
    questions. Ex vivo studies revealed that PKC activation acutely decreased DAT
    surface expression selectively in ventral, but not dorsal, striatum. AAV-mediated,
    conditional Rit2 knockdown in DAergic neurons impacted baseline DAT surface:intracellular
    distribution in DAergic terminals from female ventral, but not dorsal, striatum.
    Further, Rit2 was required for PKC-stimulated DAT internalization in both male
    and female ventral striatum. FRET and surface pulldown studies in cell lines revealed
    that PKC activation drives DAT-Rit2 surface dissociation and that the DAT N terminus
    is required for both PKC-mediated DAT-Rit2 dissociation and DAT internalization.
    Finally, we found that Rit2 and Ack1 independently converge on DAT to facilitate
    PKC-stimulated DAT endocytosis. Together, our data provide greater insight into
    mechanisms that mediate PKC-regulated DAT internalization and reveal unexpected
    region-specific differences in PKC-stimulated DAT trafficking in bona fide DAergic
    terminals. '
article_processing_charge: No
article_type: original
author:
- first_name: Rita R.
  full_name: Fagan, Rita R.
  last_name: Fagan
- first_name: Patrick J.
  full_name: Kearney, Patrick J.
  last_name: Kearney
- first_name: Carolyn G.
  full_name: Sweeney, Carolyn G.
  last_name: Sweeney
- first_name: Dino
  full_name: Luethi, Dino
  last_name: Luethi
- first_name: Florianne E
  full_name: Schoot Uiterkamp, Florianne E
  id: 3526230C-F248-11E8-B48F-1D18A9856A87
  last_name: Schoot Uiterkamp
- first_name: Klaus
  full_name: Schicker, Klaus
  last_name: Schicker
- first_name: Brian S.
  full_name: Alejandro, Brian S.
  last_name: Alejandro
- first_name: Lauren C.
  full_name: O'Connor, Lauren C.
  last_name: O'Connor
- first_name: Harald H.
  full_name: Sitte, Harald H.
  last_name: Sitte
- first_name: Haley E.
  full_name: Melikian, Haley E.
  last_name: Melikian
citation:
  ama: 'Fagan RR, Kearney PJ, Sweeney CG, et al. Dopamine transporter trafficking
    and Rit2 GTPase: Mechanism of action and in vivo impact. <i>Journal of Biological
    Chemistry</i>. 2020;295(16):5229-5244. doi:<a href="https://doi.org/10.1074/jbc.RA120.012628">10.1074/jbc.RA120.012628</a>'
  apa: 'Fagan, R. R., Kearney, P. J., Sweeney, C. G., Luethi, D., Schoot Uiterkamp,
    F. E., Schicker, K., … Melikian, H. E. (2020). Dopamine transporter trafficking
    and Rit2 GTPase: Mechanism of action and in vivo impact. <i>Journal of Biological
    Chemistry</i>. ASBMB Publications. <a href="https://doi.org/10.1074/jbc.RA120.012628">https://doi.org/10.1074/jbc.RA120.012628</a>'
  chicago: 'Fagan, Rita R., Patrick J. Kearney, Carolyn G. Sweeney, Dino Luethi, Florianne
    E Schoot Uiterkamp, Klaus Schicker, Brian S. Alejandro, Lauren C. O’Connor, Harald
    H. Sitte, and Haley E. Melikian. “Dopamine Transporter Trafficking and Rit2 GTPase:
    Mechanism of Action and in Vivo Impact.” <i>Journal of Biological Chemistry</i>.
    ASBMB Publications, 2020. <a href="https://doi.org/10.1074/jbc.RA120.012628">https://doi.org/10.1074/jbc.RA120.012628</a>.'
  ieee: 'R. R. Fagan <i>et al.</i>, “Dopamine transporter trafficking and Rit2 GTPase:
    Mechanism of action and in vivo impact,” <i>Journal of Biological Chemistry</i>,
    vol. 295, no. 16. ASBMB Publications, pp. 5229–5244, 2020.'
  ista: 'Fagan RR, Kearney PJ, Sweeney CG, Luethi D, Schoot Uiterkamp FE, Schicker
    K, Alejandro BS, O’Connor LC, Sitte HH, Melikian HE. 2020. Dopamine transporter
    trafficking and Rit2 GTPase: Mechanism of action and in vivo impact. Journal of
    Biological Chemistry. 295(16), 5229–5244.'
  mla: 'Fagan, Rita R., et al. “Dopamine Transporter Trafficking and Rit2 GTPase:
    Mechanism of Action and in Vivo Impact.” <i>Journal of Biological Chemistry</i>,
    vol. 295, no. 16, ASBMB Publications, 2020, pp. 5229–44, doi:<a href="https://doi.org/10.1074/jbc.RA120.012628">10.1074/jbc.RA120.012628</a>.'
  short: R.R. Fagan, P.J. Kearney, C.G. Sweeney, D. Luethi, F.E. Schoot Uiterkamp,
    K. Schicker, B.S. Alejandro, L.C. O’Connor, H.H. Sitte, H.E. Melikian, Journal
    of Biological Chemistry 295 (2020) 5229–5244.
date_created: 2020-05-24T22:00:59Z
date_published: 2020-04-17T00:00:00Z
date_updated: 2023-08-21T06:26:22Z
day: '17'
department:
- _id: SaSi
doi: 10.1074/jbc.RA120.012628
external_id:
  isi:
  - '000530288000006'
  pmid:
  - '32132171'
intvolume: '       295'
isi: 1
issue: '16'
language:
- iso: eng
main_file_link:
- open_access: '1'
  url: https://escholarship.umassmed.edu/oapubs/4187
month: '04'
oa: 1
oa_version: Submitted Version
page: 5229-5244
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
  eissn:
  - 1083351X
  issn:
  - '00219258'
publication_status: published
publisher: ASBMB Publications
quality_controlled: '1'
scopus_import: '1'
status: public
title: 'Dopamine transporter trafficking and Rit2 GTPase: Mechanism of action and
  in vivo impact'
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 295
year: '2020'
...