A kinome wide screen identifies novel kinases involved in regulation of monoamine transporter function

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A kinome wide screen identifies novel kinases involved in regulation of monoamine transporter function. / Vuorenpää, Anne Elina; Ammendrup-Johnsen, Ina; Jorgensen, Trine N.; Gether, Ulrik.

In: Neurochemistry International, Vol. 98, 09.2016, p. 103-114.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Vuorenpää, AE, Ammendrup-Johnsen, I, Jorgensen, TN & Gether, U 2016, 'A kinome wide screen identifies novel kinases involved in regulation of monoamine transporter function', Neurochemistry International, vol. 98, pp. 103-114. https://doi.org/10.1016/j.neuint.2016.03.013

APA

Vuorenpää, A. E., Ammendrup-Johnsen, I., Jorgensen, T. N., & Gether, U. (2016). A kinome wide screen identifies novel kinases involved in regulation of monoamine transporter function. Neurochemistry International, 98, 103-114. https://doi.org/10.1016/j.neuint.2016.03.013

Vancouver

Vuorenpää AE, Ammendrup-Johnsen I, Jorgensen TN, Gether U. A kinome wide screen identifies novel kinases involved in regulation of monoamine transporter function. Neurochemistry International. 2016 Sep;98:103-114. https://doi.org/10.1016/j.neuint.2016.03.013

Author

Vuorenpää, Anne Elina ; Ammendrup-Johnsen, Ina ; Jorgensen, Trine N. ; Gether, Ulrik. / A kinome wide screen identifies novel kinases involved in regulation of monoamine transporter function. In: Neurochemistry International. 2016 ; Vol. 98. pp. 103-114.

Bibtex

@article{4508f3b1394c4ca3ba22742825b22154,
title = "A kinome wide screen identifies novel kinases involved in regulation of monoamine transporter function",
abstract = "The high affinity transporters for the monoamine neurotransmitters, dopamine, norepinephrine, and serotonin, play a key role in controlling monoaminergic neurotransmission. It is believed that the transporters (DAT, NET and SERT, respectively) are subject to tight regulation by the cellular signaling machinery to maintain monoaminergic homeostasis. Kinases constitute a pivotal role in cellular signaling, however, the regulation of monoamine transporters by the entire ensemble of kinases is unknown. Here, we perform a whole human kinome RNA interference screen to identify novel kinases involved in regulation of monoamine transporter function and surface expression. A primary screen in HEK 293 cells stably expressing DAT or SERT with siRNAs against 573 human kinases revealed 93 kinases putatively regulating transporter function. All 93 hits, which also included kinases previously implicated in monoamine transporter regulation, such as Protein kinase B (Akt) and mitogen-activated protein kinases (MAPK), were validated with a new set of siRNAs in a secondary screen. In this screen we assessed both changes in uptake and surface expression leading to selection of 11 kinases for further evaluation in HEK 293 cells transiently expressing DAT, SERT or NET. Subsequently, three kinases; salt inducible kinase 3 (SIK3), cAMP-dependent protein kinase catalytic subunit alpha (PKA C-α) and protein kinase X-linked (PrKX); were selected for additional exploration in catecholaminergic CATH.a differentiated cells (CAD) and rat chromocytoma (PC12) cells. Whereas SIK3 likely transcriptionally regulated expression of the three transfected transporters, depletion of PKA C-α was shown to decrease SERT function. Depletion of PrKX caused decreased surface expression and function of DAT without changing protein levels, suggesting that PrKX stabilizes the transporter at the cell surface. Summarized, our data provide novel insight into kinome regulation of the monoamine transporters and identifies PrKX as a yet unappreciated possible regulator of monoamine transporter function.",
keywords = "Monoamine transporter, Uptake, Surface expression, Kinase, siRNA screen, PrKX",
author = "Vuorenp{\"a}{\"a}, {Anne Elina} and Ina Ammendrup-Johnsen and Jorgensen, {Trine N.} and Ulrik Gether",
year = "2016",
month = "9",
doi = "10.1016/j.neuint.2016.03.013",
language = "English",
volume = "98",
pages = "103--114",
journal = "Neurochemistry International",
issn = "0197-0186",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - A kinome wide screen identifies novel kinases involved in regulation of monoamine transporter function

AU - Vuorenpää, Anne Elina

AU - Ammendrup-Johnsen, Ina

AU - Jorgensen, Trine N.

AU - Gether, Ulrik

PY - 2016/9

Y1 - 2016/9

N2 - The high affinity transporters for the monoamine neurotransmitters, dopamine, norepinephrine, and serotonin, play a key role in controlling monoaminergic neurotransmission. It is believed that the transporters (DAT, NET and SERT, respectively) are subject to tight regulation by the cellular signaling machinery to maintain monoaminergic homeostasis. Kinases constitute a pivotal role in cellular signaling, however, the regulation of monoamine transporters by the entire ensemble of kinases is unknown. Here, we perform a whole human kinome RNA interference screen to identify novel kinases involved in regulation of monoamine transporter function and surface expression. A primary screen in HEK 293 cells stably expressing DAT or SERT with siRNAs against 573 human kinases revealed 93 kinases putatively regulating transporter function. All 93 hits, which also included kinases previously implicated in monoamine transporter regulation, such as Protein kinase B (Akt) and mitogen-activated protein kinases (MAPK), were validated with a new set of siRNAs in a secondary screen. In this screen we assessed both changes in uptake and surface expression leading to selection of 11 kinases for further evaluation in HEK 293 cells transiently expressing DAT, SERT or NET. Subsequently, three kinases; salt inducible kinase 3 (SIK3), cAMP-dependent protein kinase catalytic subunit alpha (PKA C-α) and protein kinase X-linked (PrKX); were selected for additional exploration in catecholaminergic CATH.a differentiated cells (CAD) and rat chromocytoma (PC12) cells. Whereas SIK3 likely transcriptionally regulated expression of the three transfected transporters, depletion of PKA C-α was shown to decrease SERT function. Depletion of PrKX caused decreased surface expression and function of DAT without changing protein levels, suggesting that PrKX stabilizes the transporter at the cell surface. Summarized, our data provide novel insight into kinome regulation of the monoamine transporters and identifies PrKX as a yet unappreciated possible regulator of monoamine transporter function.

AB - The high affinity transporters for the monoamine neurotransmitters, dopamine, norepinephrine, and serotonin, play a key role in controlling monoaminergic neurotransmission. It is believed that the transporters (DAT, NET and SERT, respectively) are subject to tight regulation by the cellular signaling machinery to maintain monoaminergic homeostasis. Kinases constitute a pivotal role in cellular signaling, however, the regulation of monoamine transporters by the entire ensemble of kinases is unknown. Here, we perform a whole human kinome RNA interference screen to identify novel kinases involved in regulation of monoamine transporter function and surface expression. A primary screen in HEK 293 cells stably expressing DAT or SERT with siRNAs against 573 human kinases revealed 93 kinases putatively regulating transporter function. All 93 hits, which also included kinases previously implicated in monoamine transporter regulation, such as Protein kinase B (Akt) and mitogen-activated protein kinases (MAPK), were validated with a new set of siRNAs in a secondary screen. In this screen we assessed both changes in uptake and surface expression leading to selection of 11 kinases for further evaluation in HEK 293 cells transiently expressing DAT, SERT or NET. Subsequently, three kinases; salt inducible kinase 3 (SIK3), cAMP-dependent protein kinase catalytic subunit alpha (PKA C-α) and protein kinase X-linked (PrKX); were selected for additional exploration in catecholaminergic CATH.a differentiated cells (CAD) and rat chromocytoma (PC12) cells. Whereas SIK3 likely transcriptionally regulated expression of the three transfected transporters, depletion of PKA C-α was shown to decrease SERT function. Depletion of PrKX caused decreased surface expression and function of DAT without changing protein levels, suggesting that PrKX stabilizes the transporter at the cell surface. Summarized, our data provide novel insight into kinome regulation of the monoamine transporters and identifies PrKX as a yet unappreciated possible regulator of monoamine transporter function.

KW - Monoamine transporter

KW - Uptake

KW - Surface expression

KW - Kinase

KW - siRNA screen

KW - PrKX

U2 - 10.1016/j.neuint.2016.03.013

DO - 10.1016/j.neuint.2016.03.013

M3 - Journal article

C2 - 27020406

VL - 98

SP - 103

EP - 114

JO - Neurochemistry International

JF - Neurochemistry International

SN - 0197-0186

ER -

ID: 167917907