Calsequestrin 1 Is an Active Partner of Stromal Interaction Molecule 2 in Skeletal Muscle

Calsequestrin 1 (CASQ1) in skeletal muscle buffers and senses Ca²⁺ in the sarcoplasmic reticulum (SR). CASQ1 also regulates store-operated Ca²⁺ entry (SOCE) by binding to stromal interaction molecule 1 (STIM1). Abnormal SOCE and/or abnormal expression or mutations in CASQ1, STIM1, or STIM2 are associated with human skeletal, cardiac, or smooth muscle diseases. However, the functional relevance of CASQ1 along with STIM2 has not been studied in any tissue, including skeletal muscle. First, in the present study, it was found by biochemical approaches that CASQ1 is bound to STIM2 via its 92 N-terminal amino acids (C1 region). Next, to examine the functional relevance of the CASQ1-STIM2 interaction in skeletal muscle, the full-length wild-type CASQ1 or the C1 region was expressed in mouse primary skeletal myotubes, and the myotubes were examined using single-myotube Ca²⁺ imaging experiments and transmission electron microscopy observations. The CASQ1-STIM2 interaction via the C1 region decreased SOCE, increased intracellular Ca²⁺ release for skeletal muscle contraction, and changed intracellular Ca²⁺ distributions (high Ca²⁺ in the SR and low Ca²⁺ in the cytosol were observed). Furthermore, the C1 region itself (which lacks Ca²⁺-buffering ability but has STIM2-binding ability) decreased the expression of Ca²⁺-related proteins (canonical-type transient receptor potential cation channel type 6 and calmodulin 1) and induced mitochondrial shape abnormalities. Therefore, in skeletal muscle, CASQ1 plays active roles in Ca²⁺ movement and distribution by interacting with STIM2 as well as Ca²⁺ sensing and buffering.