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Molecular Mechanism of TMEM16A Regulation: Role of CaMKII and PP1/PP2A.

Ayon, RJ; Hawn, MB; Aoun, J; Wiwchar, M; Forrest, AS; Cunningham, F; Singer, CA; Valencik, ML; Greenwood, IA; Leblanc, N (2019) Molecular Mechanism of TMEM16A Regulation: Role of CaMKII and PP1/PP2A. Am J Physiol Cell Physiol, 317 (6). C1093-C1106. ISSN 1522-1563 https://doi.org/10.1152/ajpcell.00059.2018
SGUL Authors: Greenwood, Iain Andrew

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Abstract

This study explored the mechanism by which Ca2+-activated Cl-channels (CaCC) encoded by the Tmem16agene are regulated by CaMKII and protein phosphatases 1 (PP1) and 2A (PP2A). Ca2+-activated Cl-currents (IClCa) were recorded from HEK-293 cells expressing mouse TMEM16A. IClCawere evoked using a pipette solution in which free Ca2+concentration ([Ca2+]i) was clamped to 500 nM, in the presence (5 mM) or absence of ATP. With 5 mM ATP, IClCadecayed to below 50% of the initial current magnitude within 10 min after seal rupture. IClCarundown seen with ATP-containing pipette solution was greatly diminished by omitting ATP. IClCarecorded after 20 min of cell dialysis with 0 ATP were more than 2-fold larger than those recorded with 5 mM ATP. Intracellular application of autocamtide-2-related inhibitory peptide (5 µM) or KN-93 (10 µM), two specific CaMKII inhibitors, produced a similar attenuation of TMEM16A rundown. In contrast, internal application of okadaic acid (30 nM) or cantharidin (100 nM), two non-selective PP1 and PP2A blockers, promoted the rundown of TMEM16A in cells dialyzed with 0 ATP. Mutating Serine 528 of TMEM16A to an Alanine led to a similar inhibition of TMEM16Arundown to that exerted by either one of the two CaMKII inhibitors tested, which was not observed for three putative CaMKII consensus sites for phosphorylation (T273, T622 and S730). Our results suggest that TMEM16A-mediated CaCCs are regulated by CaMKII and PP1/PP2A. Our data also suggest that serine 528 of TMEM16A is an important contributor to the regulation of IClCaby CaMKII.

Item Type: Article
Additional Information: Copyright © 2019, American Journal of Physiology-Cell Physiology
Keywords: Ano-1, CaMKII, Calcium-activated chloride channel, Regulation, TMEM16A, Ano-1, CaMKII, Calcium-activated chloride channel, Regulation, TMEM16A, 0601 Biochemistry And Cell Biology, 0606 Physiology, 1116 Medical Physiology, Physiology
SGUL Research Institute / Research Centre: Academic Structure > Molecular and Clinical Sciences Research Institute (MCS)
Journal or Publication Title: Am J Physiol Cell Physiol
ISSN: 1522-1563
Language: eng
Dates:
DateEvent
21 November 2019Published
28 August 2019Published Online
20 August 2019Accepted
Publisher License: Publisher's own licence
Projects:
Project IDFunderFunder ID
R01 HL075477National Institutes of Healthhttp://dx.doi.org/10.13039/100000002
R01 HL091238National Institutes of Healthhttp://dx.doi.org/10.13039/100000002
R01 HL146054National Institutes of Healthhttp://dx.doi.org/10.13039/100000002
VPRI REG 1200-121-1207University of Nevada, RenoUNSPECIFIED
PG/14/57/30992British Heart Foundationhttp://dx.doi.org/10.13039/501100000274
R01 HL127192National Institutes of Healthhttp://dx.doi.org/10.13039/100000002
NCRR 5 P20 RR15581National Center for Research Resourceshttp://dx.doi.org/10.13039/100000097
1P20GM130459National Institute of General Medical Scienceshttp://dx.doi.org/10.13039/100000057
F31 HL090023National Institutes of Healthhttp://dx.doi.org/10.13039/100000002
PubMed ID: 31461344
Go to PubMed abstract
URI: https://openaccess.sgul.ac.uk/id/eprint/111145
Publisher's version: https://doi.org/10.1152/ajpcell.00059.2018

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