title: Tespa1 protein is phosphorylated in response to store-operated calcium entry.
abstract: We previously reported that Tespa1 (thymocyte-expressed, positive selection-associated gene 1) protein expressed in lymphocytes physically interacts with IP3R (Inositol 1,4,5-trisphosphate receptor), a Ca(2+) channel protein spanning endoplasmic reticulum (ER) membrane. However, the biochemical characterization of Tespa1 protein remains unknown. In this study, we have found that Tespa1 protein was posttranslationally modified upon intracellular Ca(2+) increase in thymocytes. Through the analyses using various inhibitors, store-operated Ca(2+) entry (SOCE) was found to be an essential factor for the Tespa1 protein modification induced by T cell receptor (TCR)-stimulation. Remarkably, the Ca(2+)-dependent Tespa1 protein modification was restored by in vitro protein phosphatase treatment, indicating that this modification was due to phosphorylation. Moreover, we examined whether Ca(2+)-dependent phosphorylation of Tespa1 protein would affect the physical association between Tespa1 and IP3R proteins, revealing that physical association of these proteins is maintained regardless of the presence or absence of phosphorylation of Tespa1. In addition, KRAP protein which represents substantial amino acid sequence homology to Tespa1 was also posttranslationally phosphorylated by intracellular Ca(2+) increase in HCT116 human colon cancer cells and HEK293 human embryonic kidney cells, suggesting that common signaling mechanism(s) may contribute to the molecular modification of Tespa1 and KRAP in different cellular processes. All these results suggested a novel molecular modification of Tespa1 and the existence of the regulatory pathway that SOCE affects the Tespa1-IP3R molecular complex.
authors.affiliation: Department of Cell Biology, Faculty of Medicine, Fukuoka University, 7-45-1 Nanakuma, Fukuoka 814-0180, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Fujimoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Matsuzaki
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Tanaka
authors.forename: Senji
authors.initials: S
authors.lastname: Shirasawa
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D045325:HCT116 Cells; D057809:HEK293 Cells; D006801:Humans; D007424:Intracellular Fluid / Q000378:metabolism; D019169:Jurkat Cells; D018655:Lymphocyte Count; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation / Q000502:physiology; D011499:Protein Processing, Post-Translational; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D011948:Receptors, Antigen, T-Cell; C581428:TESPA1 protein, human; C574974:Tespa1 protein, mouse; D002118:Calcium