Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

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Standard

Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking. / Andersen, Martin Nybo; Hefting, Louise Leth; Steffensen, Annette Buur; Schmitt, Nicole; Olesen, Søren-Peter; Olsen, Jesper Velgaard; Lundby, Alicia; Rasmussen, Hanne Borger.

In: American Journal of Physiology: Cell Physiology, Vol. 309, No. 10, 15.11.2015, p. C693-C706.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Andersen, MN, Hefting, LL, Steffensen, AB, Schmitt, N, Olesen, S-P, Olsen, JV, Lundby, A & Rasmussen, HB 2015, 'Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking', American Journal of Physiology: Cell Physiology, vol. 309, no. 10, pp. C693-C706. https://doi.org/10.1152/ajpcell.00383.2014

APA

Andersen, M. N., Hefting, L. L., Steffensen, A. B., Schmitt, N., Olesen, S-P., Olsen, J. V., ... Rasmussen, H. B. (2015). Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking. American Journal of Physiology: Cell Physiology, 309(10), C693-C706. https://doi.org/10.1152/ajpcell.00383.2014

Vancouver

Andersen MN, Hefting LL, Steffensen AB, Schmitt N, Olesen S-P, Olsen JV et al. Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking. American Journal of Physiology: Cell Physiology. 2015 Nov 15;309(10):C693-C706. https://doi.org/10.1152/ajpcell.00383.2014

Author

Andersen, Martin Nybo ; Hefting, Louise Leth ; Steffensen, Annette Buur ; Schmitt, Nicole ; Olesen, Søren-Peter ; Olsen, Jesper Velgaard ; Lundby, Alicia ; Rasmussen, Hanne Borger. / Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking. In: American Journal of Physiology: Cell Physiology. 2015 ; Vol. 309, No. 10. pp. C693-C706.

Bibtex

@article{44f5ae37ac6e43c79c809d2857ffbbad,
title = "Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking",
abstract = "The potassium channel Kv7.1 plays critical physiological roles in both heart and epithelial tissues. In heart, Kv7.1 and the accessory subunit KCNE1 forms the IKs current, which is enhanced by PKA mediated phosphorylation. The observed current increase requires both phosphorylation of Kv7.1 and the presence of KCNE1. However, PKA also stimulates Kv7.1 currents in epithelial tissues, such as colon, where the channel does not co-assemble with KCNE1. Here, we demonstrate that PKA activity significantly impacts the subcellular localization of Kv7.1 in Madin Darby Canine Kidney cells. While PKA inhibition reduced the fraction of channels at the cell surface, PKA activation increased it. We show that PKA inhibition lead to intracellular accumulation of Kv7.1 in late endosomes/lysosomes. By mass spectroscopy we identified eight phosphorylated residues on Kv7.1, however, none appeared to play a role in the observed response. Instead, we found that PKA acted by regulating endocytic trafficking involving the ubiquitin ligase Nedd4-2. We show that a Nedd4-2 resistant Kv7.1-mutant displayed significantly reduced intracellular accumulation upon PKA inhibition. Similar effects were observed upon siRNA knockdown of Nedd4-2. However, although Nedd4-2 is known to regulate Kv7.1 by ubiquitylation, biochemical analyses demonstrated that PKA did not influence the amount of Nedd4-2 bound to Kv7.1 or the ubiquitylation level of the channel. This suggests that PKA influences Nedd4-2-dependent Kv7.1 transport though a different molecular mechanism. In summary, we identify a novel mechanism whereby PKA can increase Kv7.1 current levels, namely by regulating Nedd4-2 dependent Kv7.1 transport.",
author = "Andersen, {Martin Nybo} and Hefting, {Louise Leth} and Steffensen, {Annette Buur} and Nicole Schmitt and S{\o}ren-Peter Olesen and Olsen, {Jesper Velgaard} and Alicia Lundby and Rasmussen, {Hanne Borger}",
note = "Copyright {\circledC} 2015, American Journal of Physiology - Cell Physiology.",
year = "2015",
month = "11",
day = "15",
doi = "10.1152/ajpcell.00383.2014",
language = "English",
volume = "309",
pages = "C693--C706",
journal = "American Journal of Physiology: Cell Physiology",
issn = "0363-6143",
publisher = "American Physiological Society",
number = "10",

}

RIS

TY - JOUR

T1 - Protein kinase A stimulates Kv7.1 surface expression by regulating Nedd4-2-dependent endocytic trafficking

AU - Andersen, Martin Nybo

AU - Hefting, Louise Leth

AU - Steffensen, Annette Buur

AU - Schmitt, Nicole

AU - Olesen, Søren-Peter

AU - Olsen, Jesper Velgaard

AU - Lundby, Alicia

AU - Rasmussen, Hanne Borger

N1 - Copyright © 2015, American Journal of Physiology - Cell Physiology.

PY - 2015/11/15

Y1 - 2015/11/15

N2 - The potassium channel Kv7.1 plays critical physiological roles in both heart and epithelial tissues. In heart, Kv7.1 and the accessory subunit KCNE1 forms the IKs current, which is enhanced by PKA mediated phosphorylation. The observed current increase requires both phosphorylation of Kv7.1 and the presence of KCNE1. However, PKA also stimulates Kv7.1 currents in epithelial tissues, such as colon, where the channel does not co-assemble with KCNE1. Here, we demonstrate that PKA activity significantly impacts the subcellular localization of Kv7.1 in Madin Darby Canine Kidney cells. While PKA inhibition reduced the fraction of channels at the cell surface, PKA activation increased it. We show that PKA inhibition lead to intracellular accumulation of Kv7.1 in late endosomes/lysosomes. By mass spectroscopy we identified eight phosphorylated residues on Kv7.1, however, none appeared to play a role in the observed response. Instead, we found that PKA acted by regulating endocytic trafficking involving the ubiquitin ligase Nedd4-2. We show that a Nedd4-2 resistant Kv7.1-mutant displayed significantly reduced intracellular accumulation upon PKA inhibition. Similar effects were observed upon siRNA knockdown of Nedd4-2. However, although Nedd4-2 is known to regulate Kv7.1 by ubiquitylation, biochemical analyses demonstrated that PKA did not influence the amount of Nedd4-2 bound to Kv7.1 or the ubiquitylation level of the channel. This suggests that PKA influences Nedd4-2-dependent Kv7.1 transport though a different molecular mechanism. In summary, we identify a novel mechanism whereby PKA can increase Kv7.1 current levels, namely by regulating Nedd4-2 dependent Kv7.1 transport.

AB - The potassium channel Kv7.1 plays critical physiological roles in both heart and epithelial tissues. In heart, Kv7.1 and the accessory subunit KCNE1 forms the IKs current, which is enhanced by PKA mediated phosphorylation. The observed current increase requires both phosphorylation of Kv7.1 and the presence of KCNE1. However, PKA also stimulates Kv7.1 currents in epithelial tissues, such as colon, where the channel does not co-assemble with KCNE1. Here, we demonstrate that PKA activity significantly impacts the subcellular localization of Kv7.1 in Madin Darby Canine Kidney cells. While PKA inhibition reduced the fraction of channels at the cell surface, PKA activation increased it. We show that PKA inhibition lead to intracellular accumulation of Kv7.1 in late endosomes/lysosomes. By mass spectroscopy we identified eight phosphorylated residues on Kv7.1, however, none appeared to play a role in the observed response. Instead, we found that PKA acted by regulating endocytic trafficking involving the ubiquitin ligase Nedd4-2. We show that a Nedd4-2 resistant Kv7.1-mutant displayed significantly reduced intracellular accumulation upon PKA inhibition. Similar effects were observed upon siRNA knockdown of Nedd4-2. However, although Nedd4-2 is known to regulate Kv7.1 by ubiquitylation, biochemical analyses demonstrated that PKA did not influence the amount of Nedd4-2 bound to Kv7.1 or the ubiquitylation level of the channel. This suggests that PKA influences Nedd4-2-dependent Kv7.1 transport though a different molecular mechanism. In summary, we identify a novel mechanism whereby PKA can increase Kv7.1 current levels, namely by regulating Nedd4-2 dependent Kv7.1 transport.

U2 - 10.1152/ajpcell.00383.2014

DO - 10.1152/ajpcell.00383.2014

M3 - Journal article

C2 - 26405101

VL - 309

SP - C693-C706

JO - American Journal of Physiology: Cell Physiology

JF - American Journal of Physiology: Cell Physiology

SN - 0363-6143

IS - 10

ER -

ID: 145720052