PMID- 33199590
OWN - NLM
STAT- MEDLINE
DCOM- 20210125
LR  - 20240330
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 117
IP  - 48
DP  - 2020 Dec 1
TI  - Allosteric modulation of alternatively spliced Ca(2+)-activated Cl(-) channels 
      TMEM16A by PI(4,5)P(2) and CaMKII.
PG  - 30787-30798
LID - 10.1073/pnas.2014520117 [doi]
AB  - Transmembrane 16A (TMEM16A, anoctamin1), 1 of 10 TMEM16 family proteins, is a 
      Cl(-) channel activated by intracellular Ca(2+) and membrane voltage. This 
      channel is also regulated by the membrane phospholipid phosphatidylinositol 
      4,5-bisphosphate [PI(4,5)P(2)]. We find that two splice variants of TMEM16A show 
      different sensitivity to endogenous PI(4,5)P(2) degradation, where TMEM16A(ac) 
      displays higher channel activity and more current inhibition by PI(4,5)P(2) 
      depletion than TMEM16A(a). These two channel isoforms differ in the alternative 
      splicing of the c-segment (exon 13). The current amplitude and PI(4,5)P(2) 
      sensitivity of both TMEM16A(ac) and (a) are significantly strengthened by 
      decreased free cytosolic ATP and by conditions that decrease phosphorylation by 
      Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). Noise analysis suggests 
      that the augmentation of currents is due to a rise of single-channel current (i), 
      but not of channel number (N) or open probability (P(O)). Mutagenesis points to 
      arginine 486 in the first intracellular loop as a putative binding site for 
      PI(4,5)P(2), and to serine 673 in the third intracellular loop as a site for 
      regulatory channel phosphorylation that modulates the action of PI(4,5)P(2) In 
      silico simulation suggests how phosphorylation of S673 allosterically and 
      differently changes the structure of the distant PI(4,5)P(2)-binding site between 
      channel splice variants with and without the c-segment exon. In sum, our study 
      reveals the following: differential regulation of alternatively spliced 
      TMEM16A(ac) and (a) by plasma membrane PI(4,5)P(2), modification of these effects 
      by channel phosphorylation, identification of the molecular sites, and 
      mechanistic explanation by in silico simulation.
FAU - Ko, Woori
AU  - Ko W
AUID- ORCID: 0000-0001-8271-6077
AD  - Department of Brain and Cognitive Sciences, Daegu Gyeongbuk Institute of Science 
      and Technology, Daegu 42988, Republic of Korea.
FAU - Jung, Seung-Ryoung
AU  - Jung SR
AD  - Department of Physiology and Biophysics, University of Washington, Seattle, WA 
      98195.
FAU - Kim, Kwon-Woo
AU  - Kim KW
AUID- ORCID: 0000-0003-0449-2172
AD  - Department of Brain and Cognitive Sciences, Daegu Gyeongbuk Institute of Science 
      and Technology, Daegu 42988, Republic of Korea.
FAU - Yeon, Jun-Hee
AU  - Yeon JH
AD  - Department of Brain and Cognitive Sciences, Daegu Gyeongbuk Institute of Science 
      and Technology, Daegu 42988, Republic of Korea.
FAU - Park, Cheon-Gyu
AU  - Park CG
AUID- ORCID: 0000-0002-4739-1913
AD  - Department of Brain and Cognitive Sciences, Daegu Gyeongbuk Institute of Science 
      and Technology, Daegu 42988, Republic of Korea.
FAU - Nam, Joo Hyun
AU  - Nam JH
AUID- ORCID: 0000-0001-8300-6974
AD  - Department of Physiology, College of Medicine, Dongguk University, Gyeongju 
      38066, Republic of Korea.
AD  - Ion Channel Disease Research Center, College of Medicine, Dongguk University, 
      Goyang, Gyeonggi-do 10326, Republic of Korea.
FAU - Hille, Bertil
AU  - Hille B
AUID- ORCID: 0000-0002-7266-1671
AD  - Department of Physiology and Biophysics, University of Washington, Seattle, WA 
      98195.
FAU - Suh, Byung-Chang
AU  - Suh BC
AD  - Department of Brain and Cognitive Sciences, Daegu Gyeongbuk Institute of Science 
      and Technology, Daegu 42988, Republic of Korea; bcsuh@dgist.ac.kr.
LA  - eng
GR  - R37 NS008174/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20201116
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (Anoctamin-1)
RN  - 0 (Phosphatidylinositols)
RN  - 0 (Protein Isoforms)
RN  - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinase Type 2)
SB  - IM
MH  - Allosteric Regulation
MH  - *Alternative Splicing
MH  - Animals
MH  - Anoctamin-1/chemistry/*genetics/*metabolism
MH  - Binding Sites
MH  - Calcium-Calmodulin-Dependent Protein Kinase Type 2/*metabolism
MH  - Cell Membrane/metabolism
MH  - Gene Expression Regulation
MH  - HEK293 Cells
MH  - Humans
MH  - Ion Channel Gating/drug effects
MH  - Mice
MH  - Models, Molecular
MH  - Molecular Conformation
MH  - Mutagenesis, Site-Directed
MH  - Phosphatidylinositols/*metabolism
MH  - Phosphorylation
MH  - Protein Binding
MH  - Protein Isoforms
MH  - Structure-Activity Relationship
PMC - PMC7720229
OTO - NOTNLM
OT  - Ca2+-activated Cl- channel
OT  - PI(4,5)P2
OT  - TMEM16A
OT  - intracellular ATP
OT  - splice variants
COIS- The authors declare no competing interest.
EDAT- 2020/11/18 06:00
MHDA- 2021/01/26 06:00
PMCR- 2021/05/16
CRDT- 2020/11/17 06:14
PHST- 2020/11/18 06:00 [pubmed]
PHST- 2021/01/26 06:00 [medline]
PHST- 2020/11/17 06:14 [entrez]
PHST- 2021/05/16 00:00 [pmc-release]
AID - 2014520117 [pii]
AID - 202014520 [pii]
AID - 10.1073/pnas.2014520117 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2020 Dec 1;117(48):30787-30798. doi: 
      10.1073/pnas.2014520117. Epub 2020 Nov 16.