Super-Resolution Microscopy Reveals That Stromal Interaction Molecule 1 Trafficking Depends on Microtubule Dynamics

Yi-Ting Huang; Ya-Ting Hsu; Yih-Fung Chen; Meng-Ru Shen

doi:10.3389/fphys.2021.762387

Super-Resolution Microscopy Reveals That Stromal Interaction Molecule 1 Trafficking Depends on Microtubule Dynamics

Front Physiol. 2021 Nov 5:12:762387. doi: 10.3389/fphys.2021.762387. eCollection 2021.

Authors

Yi-Ting Huang^{1

2}, Ya-Ting Hsu^{3

4}, Yih-Fung Chen^{5

6}, Meng-Ru Shen^{1

7}

Affiliations

¹ Department of Pharmacology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
² Institute of Basic Medical Sciences, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
³ Institute of Clinical Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
⁴ Division of Hematology, Department of Internal Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
⁵ Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan.
⁶ Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
⁷ Department of Obstetrics and Gynecology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.

Abstract

Store-operated Ca²⁺ entry (SOCE) is an essential pathway for Ca²⁺ signaling, and regulates various vital cellular functions. It is triggered by the endoplasmic reticulum Ca²⁺ sensor stromal interaction molecule 1 (STIM1). Illustration of STIM1 spatiotemporal structure at the nanometer scale during SOCE activation provides structural and functional insights into the fundamental Ca²⁺ homeostasis. In this study, we used direct stochastic optical reconstruction microscopy (dSTORM) to revisit the dynamic process of the interaction between STIM1, end-binding protein (EB), and microtubules to the ER-plasma membrane. Using dSTORM, we found that"powder-like"STIM1 aggregates into "trabecular-like" architectures toward the cell periphery during SOCE, and that an intact microtubule network and EB1 are essential for STIM1 trafficking. After thapsigargin treatment, STIM1 can interact with EB1 regardless of undergoing aggregation. We generated STIM1 variants adapted from a real-world database and introduced them into SiHa cells to clarify the impact of STIM1 mutations on cancer cell behavior. The p.D76G and p.D84Y variants locating on the Ca²⁺ binding domain of STIM1 result in inhibition of focal adhesion turnover, Ca²⁺ influx during SOCE and subsequent cell migration. Inversely, the p.R643C variant on the microtubule interacting domain of STIM1 leads to dissimilar consequence and aggravates cell migration. These findings imply that STIM1 mutational patterns have an impact on cancer metastasis, and therefore could be either a prognostic marker or a novel therapeutic target to inhibit the malignant behavior of STIM1-mediated cancer cells. Altogether, we generated novel insight into the role of STIM1 during SOCE activation, and uncovered the impact of real-world STIM1 variants on cancer cells.

Keywords: direct stochastic optical reconstruction microscopy (dSTORM); microtubule network; somatic mutation; store-operated Ca2+ entry (SOCE); stromal interaction molecule 1 (STIM1).