TY - JOUR
T1 - SNAP23 regulates KCC2 membrane insertion and activity following mZnR/GPR39 activation in hippocampalneurons
AU - Asraf, Hila
AU - Bogdanovic, Milos
AU - Gottesman, Noa
AU - Sekler, Israel
AU - Aizenman, Elias
AU - Hershfinkel, Michal
N1 - Funding Information: We thank Dr. Igor Medina for critical reading of the manuscript and his advice and support throughout this project. This work was funded by a joint National Science Foundation-US-Israel Binational Science Foundation grant NSF-IOS-BSF 1655480 (MH, EA). MH is also funded by Israel Science Foundation grant 812/20. H.A and M.H designed experiments, H. A. N.G. and M.B. performed experiments and analyzed data, H.A. I.S. E. A. and M.H. interpreted the data, wrote and edited the manuscript. The authors declare no competing interests. Funding Information: We thank Dr. Igor Medina for critical reading of the manuscript and his advice and support throughout this project. This work was funded by a joint National Science Foundation -US-Israel Binational Science Foundation grant NSF-IOS-BSF 1655480 (MH, EA). MH is also funded by Israel Science Foundation grant 812/20. Publisher Copyright: © 2022 The Authors
PY - 2022/2/18
Y1 - 2022/2/18
N2 - Modulation of the neuronal K+/Cl− cotransporter 2 (KCC2) activity, which mediates Cl− export, is critical to neuronal function. Here, we demonstrate that KCC2 interacts with the SNARE protein synaptosome-associated protein 23, SNAP23, an essential component of membrane insertion machinery. Using KCC2 truncated mutants, we show that KCC2 C-terminal domain is essential for membrane targeting and SNAP23-dependent upregulation of KCC2 activity triggered by activation of the Zn2+-sensitive receptor mZnR/GPR39 in HEK293 cells. Expression of SNAP23 phosphorylation-insensitive mutants or inhibition of its upstream activator IκB kinase (IKK) prevents mZnR/GPR39 upregulation of KCC2 activity in mouse hippocampal neurons. We further find that SNAP23 interacts with Syntaxin 1A and KCC2, and that all three proteins exhibit increased membrane insertion following mZnR/GPR39 activation in neurons. Our results elucidate a G-protein-coupled receptor-dependent pathway for regulation of KCC activity, mediated via interaction with SNARE proteins.
AB - Modulation of the neuronal K+/Cl− cotransporter 2 (KCC2) activity, which mediates Cl− export, is critical to neuronal function. Here, we demonstrate that KCC2 interacts with the SNARE protein synaptosome-associated protein 23, SNAP23, an essential component of membrane insertion machinery. Using KCC2 truncated mutants, we show that KCC2 C-terminal domain is essential for membrane targeting and SNAP23-dependent upregulation of KCC2 activity triggered by activation of the Zn2+-sensitive receptor mZnR/GPR39 in HEK293 cells. Expression of SNAP23 phosphorylation-insensitive mutants or inhibition of its upstream activator IκB kinase (IKK) prevents mZnR/GPR39 upregulation of KCC2 activity in mouse hippocampal neurons. We further find that SNAP23 interacts with Syntaxin 1A and KCC2, and that all three proteins exhibit increased membrane insertion following mZnR/GPR39 activation in neurons. Our results elucidate a G-protein-coupled receptor-dependent pathway for regulation of KCC activity, mediated via interaction with SNARE proteins.
KW - Cell biology
KW - Molecular physiology
KW - Neuroscience
UR - http://www.scopus.com/inward/record.url?scp=85123708775&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.isci.2022.103751
DO - https://doi.org/10.1016/j.isci.2022.103751
M3 - Article
C2 - 35118363
SN - 2589-0042
VL - 25
JO - iScience
JF - iScience
IS - 2
M1 - 103751
ER -