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ER-localized bestrophin 1 activates Ca2+-dependent ion channels TMEM16A and SK4 possibly by acting as a counterion channel

  • Signaling and Cell Physiology
  • Published:
Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

Bestrophins form Ca2+-activated Cl− channels and regulate intracellular Ca2+ signaling. We demonstrate that bestrophin 1 is localized in the endoplasmic reticulum (ER), where it interacts with stromal interacting molecule 1, the ER-Ca2+ sensor. Intracellular Ca2+ transients elicited by stimulation of purinergic P2Y2 receptors in HEK293 cells were augmented by hBest1. The p21-activated protein kinase Pak2 was found to phosphorylate hBest1, thereby enhancing Ca2+ signaling and activation of Ca2+-dependent Cl− (TMEM16A) and K+ (SK4) channels. Lack of bestrophin 1 expression in respiratory epithelial cells of mBest1 knockout mice caused expansion of ER cisterns and induced Ca2+ deposits. hBest1 is, therefore, important for Ca2+ handling of the ER store and may resemble the long-suspected counterion channel to balance transient membrane potentials occurring through inositol triphosphate (IP3)-induced Ca2+ release and store refill. Thus, bestrophin 1 regulates compartmentalized Ca2+ signaling that plays an essential role in Best macular dystrophy, inflammatory diseases such as cystic fibrosis, as well as proliferation.

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Abbreviations

hBest1:

human bestrophin 1

CaCC:

Ca2+-activated Cl− channels

SK4:

small conductance calcium-activated potassium channel type 4

TMEM16A:

transmembrane protein 16A

ANO1:

anoctamin 1

Pak2:

p21-activated protein kinase

ER:

endoplasmic reticulum

SERCA:

sarcoendoplasmic reticulum calcium ATPase

Stim1:

stromal interacting molecule 1

DIDS:

4,4′-diisothio-cyanostilbene-2,2′-disulfonic acid

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Acknowledgments

This study was supported by the grants DFG SFB699 A6/A7, DFG KU 756/8-2, and Else Kröner-Fresenius-Stiftung P36/05//A44/05. We are grateful for the technical expertise of Caio Toledo, Christine Meese, Karin Schadendorf, Helga Schmidt, and Uwe de Vries in performing the ultrastructural analysis. The Ca2+-sensitive GFP protein G-CaMP2 was kindly provided by Dr. J. Nakai, Wako City Saitama, Japan. We gratefully acknowledge the supply of the vmd2−/− mice by MERCK Research Laboratories (770 Sumneytown Pike, West Point, PA, USA).

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Authors and Affiliations

  1. Institut für Physiologie, Universität Regensburg, Universitätsstraße 31, 93053, Regensburg, Germany

    René Barro-Soria, Fadi Aldehni, Joana Almaça, Rainer Schreiber & Karl Kunzelmann

  2. Institut für Anatomie, Universität Regensburg, Universitätsstraße 31, 93053, Regensburg, Germany

    Ralph Witzgall

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  1. René Barro-Soria
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  2. Fadi Aldehni
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  3. Joana Almaça
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  6. Karl Kunzelmann
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Correspondence to Karl Kunzelmann.

Additional information

René Barro-Soria and Fadi Aldehni contributed equally to the present study.

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Barro-Soria, R., Aldehni, F., Almaça, J. et al. ER-localized bestrophin 1 activates Ca2+-dependent ion channels TMEM16A and SK4 possibly by acting as a counterion channel. Pflugers Arch - Eur J Physiol 459, 485–497 (2010). https://doi.org/10.1007/s00424-009-0745-0

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