論文

2020年10月6日

Fluctuation in O-GlcNAcylation inactivates STIM1 to reduce store-operated calcium ion entry via downregulation of Ser621 phosphorylation

Journal of Biological Chemistry
  • Atsuo Nomura
  • ,
  • Shunichi Yokoe
  • ,
  • Kiichiro Tomoda
  • ,
  • Takatoshi Nakagawa
  • ,
  • Francisco Javier Martin-Romero
  • ,
  • Michio Asahi

開始ページ
jbc.RA120.014271
終了ページ
jbc.RA120.014271
記述言語
掲載種別
研究論文(学術雑誌)
DOI
10.1074/jbc.ra120.014271
出版者・発行元
American Society for Biochemistry & Molecular Biology (ASBMB)

Stromal interaction molecule 1 (STIM1) plays a pivotal role in store-operated Ca2+ entry (SOCE), an essential mechanism in cellular calcium signaling and in maintaining cellular calcium balance. Because <italic>O</italic>-GlcNAcylation plays pivotal roles in various cellular function, we examined the effect of fluctuation in STIM1 <italic>O</italic>-GlcNAcylation on SOCE activity. We found that both increase and decrease in STIM1 <italic>O</italic>-GlcNAcylation impaired SOCE activity. To determine the molecular basis, we established STIM1-knockout HEK293 (STIM1-KO-HEK) cells using the CRISPR/Cas9 system and transfected STIM1 wild-type (STIM1-KO-WT-HEK), S621A (STIM1-KO-S621A-HEK), or T626A (STIM1-KO-T626A-HEK) cells. Using these cells, we examined the possible <italic>O</italic>-GlcNAcylation sites of STIM1 to determine whether the sites were <italic>O</italic>-GlcNAcylated. Co-immunoprecipitation analysis revealed that Ser621 and Thr626 were <italic>O</italic>-GlcNAcylated and that Thr626 was <italic>O</italic>-GlcNAcylated in the steady state but Ser621 was not. The SOCE activity in STIM1-KO-S621A-HEK and STIM1-KO-T626A-HEK cells was lower than that in STIM1-KO-WT-HEK cells because of reduced phosphorylation at Ser621. Treatment with the <italic>O</italic>-GlcNAcase inhibitor Thiamet G or <italic>O</italic>-GlcNAc transferase (OGT) transfection, which increases <italic>O</italic>-GlcNAcylation, reduced SOCE activity, whereas treatment with the OGT inhibitor ST045849 or siOGT transfection, which decreases <italic>O</italic>-GlcNAcylation, also reduced SOCE activity. Decrease in SOCE activity due to increase and decrease in <italic>O</italic>-GlcNAcylation was attributable to reduced phosphorylation at Ser621. These data suggest that both decrease in <italic>O</italic>-GlcNAcylation at Thr626 and increase in<italic> O</italic>-GlcNAcylation at Ser621 in STIM1 lead to impairment of SOCE activity through decrease in Ser621 phosphorylation. Targeting STIM1 <italic>O</italic>-GlcNAcylation could provide a promising treatment option for the related diseases such as neurodegenerative diseases.

リンク情報
DOI
https://doi.org/10.1074/jbc.ra120.014271
URL
https://syndication.highwire.org/content/doi/10.1074/jbc.RA120.014271
ID情報
  • DOI : 10.1074/jbc.ra120.014271
  • ISSN : 0021-9258
  • eISSN : 1083-351X

エクスポート
BibTeX RIS