Mapping the interacting domains of STIM1 and Orai1 in Ca2+ release-activated Ca2+ channel activation.
STIM1 and Orai1 are essential components of Ca(2+) release-activated Ca(2+) channels (CRACs). After endoplasmic reticulum Ca(2+) store depletion, STIM1 in the endoplasmic reticulum aggregates and migrates toward the cell periphery to co-localize with Orai1 on the opposing plasma membrane. Little is known about the roles of different domains of STIM1 and Orai1 in protein clustering, migration, interaction, and, ultimately, opening CRAC channels. Here we demonstrate that the coiled-coil domain in the C terminus of STIM1 is crucial for its aggregation. Amino acids 425-671 of STIM1, which contain a serine-proline-rich region, are important for the correct targeting of the STIM1 cluster to the cell periphery after calcium store depletion. The polycationic region in the C-terminal tail of STIM1 also helps STIM1 targeting but is not essential for CRAC channel activation. The cytoplasmic C terminus but not the N terminus of Orai1 is required for its interaction with STIM1. We further identify a highly conserved region in the N terminus of Orai1 (amino acids 74-90) that is necessary for CRAC channel opening. Finally, we show that the transmembrane domain of Orai1 participates in Orai1-Orai1 interactions.
Li
Zhengzheng
Z
National Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Datun Road 15, Chaoyang District, Beijing 100101, China.
Lu
Jingze
J
Xu
Pingyong
P
Xie
Xiangyang
X
Chen
Liangyi
L
Xu
Tao
T
D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000737:chemistry*; D002412:Cations; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D004594:Electrophysiology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*; D009154:Mutation; D009363:Neoplasm Proteins / Q000737:chemistry*; D000071740:ORAI1 Protein; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D000071737:Stromal Interaction Molecule 1; D013997:Time Factors; D014162:Transfection
D015220:Calcium Channels; D002412:Cations; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D000071740:ORAI1 Protein; C510746:ORAI1 protein, human; C103530:STIM1 protein, human; D000071737:Stromal Interaction Molecule 1; D002118:Calcium