PMID- 35805121
OWN - NLM
STAT- MEDLINE
DCOM- 20220712
LR  - 20220828
IS  - 2073-4409 (Electronic)
IS  - 2073-4409 (Linking)
VI  - 11
IP  - 13
DP  - 2022 Jun 27
TI  - PKC-Mediated Orai1 Channel Phosphorylation Modulates Ca(2+) Signaling in HeLa 
      Cells.
LID - 10.3390/cells11132037 [doi]
LID - 2037
AB  - The overexpression of the Orai1 channel inhibits SOCE when using the Ca(2+) 
      readdition protocol. However, we found that HeLa cells overexpressing the Orai1 
      channel displayed enhanced Ca(2+) entry and a limited ER depletion in response to 
      the combination of ATP and thapsigargin (TG) in the presence of external Ca(2+). 
      As these effects require the combination of an agonist and TG, we decided to 
      study whether the phosphorylation of Orai1 S27/S30 residues had any role using 
      two different mutants: Orai1-S27/30A (O1-AA, phosphorylation-resistant) and 
      Orai1-S27/30D (O1-DD, phosphomimetic). Both O1-wt and O1-AA supported enhanced 
      Ca(2+) entry, but this was not the case with O1-E106A (dead-pore mutant), O1-DD, 
      and O1-AA-E106A, while O1-wt, O1-E106A, and O1-DD inhibited the ATP and 
      TG-induced reduction of ER [Ca(2+)], suggesting that the phosphorylation of O1 
      S27/30 interferes with the IP(3)R activity. O1-wt and O1-DD displayed an 
      increased interaction with IP(3)R in response to ATP and TG; however, the O1-AA 
      channel decreased this interaction. The expression of mCherry-O1-AA increased the 
      frequency of ATP-induced sinusoidal [Ca(2+)](i) oscillations, while mCherry-O1-wt 
      and mCherry-O1-DD decreased this frequency. These data suggest that the 
      combination of ATP and TG stimulates Ca(2+) entry, and the phosphorylation of 
      Orai1 S27/30 residues by PKC reduces IP(3)R-mediated Ca(2+) release.
FAU - Martinez-Martinez, Ericka
AU  - Martinez-Martinez E
AD  - Department of Biochemistry, Cinvestav, Mexico City 07360, Mexico.
FAU - Sanchez-Vazquez, Victor Hugo
AU  - Sanchez-Vazquez VH
AD  - Department of Biochemistry, Cinvestav, Mexico City 07360, Mexico.
FAU - Leon-Aparicio, Daniel
AU  - Leon-Aparicio D
AD  - Department of Biochemistry, Cinvestav, Mexico City 07360, Mexico.
AD  - Instituto de Fisica, Universidad Autonoma de San Luis Potosi, San Luis Potosi 
      78290, Mexico.
FAU - Sanchez-Collado, Jose
AU  - Sanchez-Collado J
AD  - Cellular Physiology Research Group, Department of Physiology, Institute of 
      Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain.
FAU - Gallegos-Gomez, Martin-Leonardo
AU  - Gallegos-Gomez ML
AD  - Department of Biochemistry, Cinvestav, Mexico City 07360, Mexico.
FAU - Rosado, Juan A
AU  - Rosado JA
AUID- ORCID: 0000-0002-9749-2325
AD  - Cellular Physiology Research Group, Department of Physiology, Institute of 
      Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain.
FAU - Arias, Juan M
AU  - Arias JM
AD  - Programa de Neurociencias-UIICSE, Facultad de Estudios Superiores Iztacala, 
      Universidad Nacional Autonoma de Mexico, Tlalnepantla de Baz 54090, Mexico.
FAU - Guerrero-Hernandez, Agustin
AU  - Guerrero-Hernandez A
AUID- ORCID: 0000-0002-8653-4841
AD  - Department of Biochemistry, Cinvestav, Mexico City 07360, Mexico.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20220627
PL  - Switzerland
TA  - Cells
JT  - Cells
JID - 101600052
RN  - 0 (Calcium Channels)
RN  - 0 (ORAI1 Protein)
RN  - 0 (ORAI1 protein, human)
RN  - 67526-95-8 (Thapsigargin)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 2.7.11.13 (Protein Kinase C)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Adenosine Triphosphate/metabolism/pharmacology
MH  - *Calcium/metabolism
MH  - *Calcium Channels/metabolism
MH  - HeLa Cells
MH  - Humans
MH  - ORAI1 Protein/metabolism
MH  - Phosphorylation
MH  - Protein Kinase C/metabolism
MH  - Thapsigargin/pharmacology
PMC - PMC9266177
OTO - NOTNLM
OT  - PKC
OT  - SOCE
OT  - phosphomimetic
OT  - phosphorylated Orai1
COIS- The authors declare no conflict of interest.
EDAT- 2022/07/10 06:00
MHDA- 2022/07/14 06:00
PMCR- 2022/06/27
CRDT- 2022/07/09 01:05
PHST- 2022/03/17 00:00 [received]
PHST- 2022/05/15 00:00 [revised]
PHST- 2022/05/27 00:00 [accepted]
PHST- 2022/07/09 01:05 [entrez]
PHST- 2022/07/10 06:00 [pubmed]
PHST- 2022/07/14 06:00 [medline]
PHST- 2022/06/27 00:00 [pmc-release]
AID - cells11132037 [pii]
AID - cells-11-02037 [pii]
AID - 10.3390/cells11132037 [doi]
PST - epublish
SO  - Cells. 2022 Jun 27;11(13):2037. doi: 10.3390/cells11132037.