title: STIM1 negatively regulates Ca²⁺ release from the sarcoplasmic reticulum in skeletal myotubes.
abstract: STIM1 (stromal interaction molecule 1) mediates SOCE (store-operated Ca²⁺ entry) in skeletal muscle. However, the direct role(s) of STIM1 in skeletal muscle, such as Ca²⁺ release from the SR (sarcoplasmic reticulum) for muscle contraction, have not been identified. The times required for the maximal expression of endogenous STIM1 or Orai1, or for the appearance of puncta during the differentiation of mouse primary skeletal myoblasts to myotubes, were all different, and the formation of puncta was detected with no stimulus during differentiation, suggesting that, in skeletal muscle, the formation of puncta is a part of the differentiation. Wild-type STIM1 and two STIM1 mutants (Triple mutant, missing Ca²⁺-sensing residues but possessing the intact C-terminus; and E136X, missing the C-terminus) were overexpressed in the myotubes. The wild-type STIM1 increased SOCE, whereas neither mutant had an effect on SOCE. It was interesting that increases in the formation of puncta were observed in the Triple mutant as well as in wild-type STIM1, suggesting that SOCE-irrelevant puncta could exist in skeletal muscle. On the other hand, overexpression of wild-type or Triple mutant, but not E136X, attenuated Ca²⁺ releases from the SR in response to KCl [evoking ECC (excitation-contraction coupling) via activating DHPR (dihydropyridine receptor)] in a dominant-negative manner. The attenuation was removed by STIM1 knockdown, and STIM1 was co-immunoprecipitated with DHRP in a Ca²⁺-independent manner. These results suggest that STIM1 negatively regulates Ca²⁺ release from the SR through the direct interaction of the STIM1 C-terminus with DHPR, and that STIM1 is involved in both ECC and SOCE in skeletal muscle.
authors.affiliation: *Department of Physiology, College of Medicine, The Catholic University of Korea, Seoul 137-701, Korea.
authors.forename: Keon Jin
authors.initials: KJ
authors.lastname: Lee
authors.forename: Jin Seok
authors.initials: JS
authors.lastname: Woo
authors.forename: Ji-Hye
authors.initials: JH
authors.lastname: Hwang
authors.forename: Changdo
authors.initials: C
authors.lastname: Hyun
authors.forename: Chung-Hyun
authors.initials: CH
authors.lastname: Cho
authors.forename: Do Han
authors.initials: DH
authors.lastname: Kim
authors.forename: Eun Hui
authors.initials: EH
authors.lastname: Lee
mesh_terms: D000818:Animals; D001483:Base Sequence; D001842:Bone and Bones / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D017931:DNA Primers; D018076:DNA, Complementary; D006801:Humans; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*; D000071737:Stromal Interaction Molecule 1
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D019837:Ryanodine Receptor Calcium Release Channel; C103530:STIM1 protein, human; D000071737:Stromal Interaction Molecule 1; D002118:Calcium