Structural, functional, and behavioral insights of dopamine dysfunction revealed by a deletion in SLC6A3

Nicholas G. Campbell, Aparna Shekar, Jenny I. Aguilar, Dungeng Peng, Vikas Navratna, Dongxue Yang, Alexander N. Morley, Amanda M. Duran, Greta Galli, Brian O’Grady, Ramnarayan Ramachandran, James S. Sutcliffe, Harald H. Sitte, Kevin Erreger, Jens Meiler, Thomas Stockner, Leon M. Bellan, Heinrich J.G. Matthies, Eric Gouaux, Hassane S. MchaourabAurelio Galli

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The human dopamine (DA) transporter (hDAT) mediates clearance of DA. Genetic variants in hDAT have been associated with DA dysfunction, a complication associated with several brain disorders, including autism spectrum disorder (ASD). Here, we investigated the structural and behavioral bases of an ASD-associated in-frame deletion in hDAT at N336 (ΔN336). We uncovered that the deletion promoted a previously unobserved conformation of the intracellular gate of the transporter, likely representing the rate-limiting step of the transport process. It is defined by a “half-open and inward-facing” state (HOIF) of the intracellular gate that is stabilized by a network of interactions conserved phylogenetically, as we demonstrated in hDAT by Rosetta molecular modeling and fine-grained simulations, as well as in its bacterial homolog leucine transporter by electron paramagnetic resonance analysis and X-ray crystallography. The stabilization of the HOIF state is associated both with DA dysfunctions demonstrated in isolated brains of Drosophila melanogaster expressing hDAT ΔN336 and with abnormal behaviors observed at high-time resolution. These flies display increased fear, impaired social interactions, and locomotion traits we associate with DA dysfunction and the HOIF state. Together, our results describe how a genetic variation causes DA dysfunction and abnormal behaviors by stabilizing a HOIF state of the transporter.

Original languageEnglish (US)
Pages (from-to)3853-3862
Number of pages10
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number9
DOIs
StatePublished - Feb 26 2019

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Dopamine
Dopamine Plasma Membrane Transport Proteins
X Ray Crystallography
Electron Spin Resonance Spectroscopy
Brain Diseases
Locomotion
Interpersonal Relations
Drosophila melanogaster
Leucine
Diptera
Fear
Brain
Autism Spectrum Disorder

Keywords

  • Amphetamine
  • Autism
  • Dopamine transporter
  • Efflux
  • Leucine transporter

ASJC Scopus subject areas

  • General

Cite this

Structural, functional, and behavioral insights of dopamine dysfunction revealed by a deletion in SLC6A3. / Campbell, Nicholas G.; Shekar, Aparna; Aguilar, Jenny I.; Peng, Dungeng; Navratna, Vikas; Yang, Dongxue; Morley, Alexander N.; Duran, Amanda M.; Galli, Greta; O’Grady, Brian; Ramachandran, Ramnarayan; Sutcliffe, James S.; Sitte, Harald H.; Erreger, Kevin; Meiler, Jens; Stockner, Thomas; Bellan, Leon M.; Matthies, Heinrich J.G.; Gouaux, Eric; Mchaourab, Hassane S.; Galli, Aurelio.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 9, 26.02.2019, p. 3853-3862.

Research output: Contribution to journalArticle

Campbell, NG, Shekar, A, Aguilar, JI, Peng, D, Navratna, V, Yang, D, Morley, AN, Duran, AM, Galli, G, O’Grady, B, Ramachandran, R, Sutcliffe, JS, Sitte, HH, Erreger, K, Meiler, J, Stockner, T, Bellan, LM, Matthies, HJG, Gouaux, E, Mchaourab, HS & Galli, A 2019, 'Structural, functional, and behavioral insights of dopamine dysfunction revealed by a deletion in SLC6A3', Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 9, pp. 3853-3862. https://doi.org/10.1073/pnas.1816247116
Campbell, Nicholas G. ; Shekar, Aparna ; Aguilar, Jenny I. ; Peng, Dungeng ; Navratna, Vikas ; Yang, Dongxue ; Morley, Alexander N. ; Duran, Amanda M. ; Galli, Greta ; O’Grady, Brian ; Ramachandran, Ramnarayan ; Sutcliffe, James S. ; Sitte, Harald H. ; Erreger, Kevin ; Meiler, Jens ; Stockner, Thomas ; Bellan, Leon M. ; Matthies, Heinrich J.G. ; Gouaux, Eric ; Mchaourab, Hassane S. ; Galli, Aurelio. / Structural, functional, and behavioral insights of dopamine dysfunction revealed by a deletion in SLC6A3. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 9. pp. 3853-3862.
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abstract = "The human dopamine (DA) transporter (hDAT) mediates clearance of DA. Genetic variants in hDAT have been associated with DA dysfunction, a complication associated with several brain disorders, including autism spectrum disorder (ASD). Here, we investigated the structural and behavioral bases of an ASD-associated in-frame deletion in hDAT at N336 (ΔN336). We uncovered that the deletion promoted a previously unobserved conformation of the intracellular gate of the transporter, likely representing the rate-limiting step of the transport process. It is defined by a “half-open and inward-facing” state (HOIF) of the intracellular gate that is stabilized by a network of interactions conserved phylogenetically, as we demonstrated in hDAT by Rosetta molecular modeling and fine-grained simulations, as well as in its bacterial homolog leucine transporter by electron paramagnetic resonance analysis and X-ray crystallography. The stabilization of the HOIF state is associated both with DA dysfunctions demonstrated in isolated brains of Drosophila melanogaster expressing hDAT ΔN336 and with abnormal behaviors observed at high-time resolution. These flies display increased fear, impaired social interactions, and locomotion traits we associate with DA dysfunction and the HOIF state. Together, our results describe how a genetic variation causes DA dysfunction and abnormal behaviors by stabilizing a HOIF state of the transporter.",
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AU - Navratna, Vikas

AU - Yang, Dongxue

AU - Morley, Alexander N.

AU - Duran, Amanda M.

AU - Galli, Greta

AU - O’Grady, Brian

AU - Ramachandran, Ramnarayan

AU - Sutcliffe, James S.

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AU - Erreger, Kevin

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