Esterase D stabilizes FKBP25 to suppress mTORC1

Cellular & Molecular Biology Letters

ZhaoMin Lin¹  Baoxiang Zhao²  Na Li³  Junying Miao³  Yuejun Yang³  Wen Yao³  Xiaoling Cui³  Xinpeng Chen⁴ 

[1] Institute of Medical Science, The Second Hospital of Shandong University, 250033, Jinan, People’s Republic of China;Institute of Organic Chemistry, School of Chemistry and Chemical Engineering, Shandong University, 250100, Jinan, People’s Republic of China;Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Science, Shandong University, 266237, Qingdao, People’s Republic of China;Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Science, Shandong University, 266237, Qingdao, People’s Republic of China;Hubei Key Laboratory of Edible Wild Plants Conservation & Utilization, Hubei Engineering Research Center of Typical Wild Vegetable Breeding and Comprehensive Utilization Technology, Hubei Normal University, 435002, Huangshi, People’s Republic of China;

关键词: Esterase D;    FKBP25;    mTORC1;    Ubiquitination;    Autophagy;   

DOI  :  10.1186/s11658-021-00297-2

来源: Springer

【 摘 要 】

BackgroundEsterase D (ESD) is a nonspecific esterase that detoxifies formaldehyde. Many reports have stated that ESD activity is associated with a variety of physiological and pathological processes. However, the detailed signaling pathway of ESD remains poorly understood.MethodsConsidering the advantages of the small chemical molecule, our recent work demonstrated that 4-chloro-2-(5-phenyl-1-(pyridin-2-yl)-4,5-dihydro-1H-pyrazol-3-yl) phenol (FPD5) activates ESD, and will be a good tool for studying ESD further. Firstly, we determined the interaction between ESD and FK506 binding protein 25 (FKBP25) by yeast two-hybrid assay and co-immunoprecipitation (CO-IP) and analyzed the phosphorylation levels of mTORC1, P70S6K and 4EBP1 by western blot. Furthermore, we used the sulforhodamine B (SRB) and chick chorioallantoic membrane (CAM) assay to analyze cell viability in vitro and in vivo after treatment with ESD activator FPD5.ResultsWe screened FKBP25 as a candidate protein to interact with ESD by yeast two-hybrid assay. Then we verified the interaction between ESD and endogenous FKBP25 or ectopically expressed GFP-FKBP25 by CO-IP. Moreover, the N-terminus (1–90 aa) domain of FKBP25 served as the crucial element for their interaction. More importantly, ESD reduced the K48-linked poly-ubiquitin chains of FKBP25 and thus stabilized cytoplasmic FKBP25. ESD also promoted FKBP25 to bind more mTORC1, suppressing the activity of mTORC1. In addition, ESD suppressed tumor cell growth in vitro and in vivo through autophagy.ConclusionsThese findings provide novel evidence for elucidating the molecular mechanism of ESD and ubiquitination of FKBP25 to regulate autophagy and cancer cell growth. The ESD/FKBP25/mTORC1 signaling pathway is involved in inhibiting tumor cell growth via regulating autophagy.

【 授权许可】

CC BY