Esterase D stabilizes FKBP25 to suppress mTORC1.

Abstract

BACKGROUND:

Esterase 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.

METHODS:

Considering 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.

RESULTS:

We 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.

CONCLUSIONS:

These 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.

• Authors: Yuejun Yang, Xinpeng Chen, Wen Yao, Xiaoling Cui, Na Li, ZhaoMin Lin, Baoxiang Zhao, Junying Miao
• Journal: Cellular & molecular biology letters (Cell Mol Biol Lett) Vol. 26 Issue 1 Pg. 50 (Dec 07 2021) ISSN: 1689-1392 [Electronic] England
• PMID: 34875997 (Publication Type: Journal Article)
• Copyright: © 2021. The Author(s).

Chemical References

• Pyrazoles
• FKBP3 protein, human
• pyrazole
• Mechanistic Target of Rapamycin Complex 1
• Thiolester Hydrolases
• s-formylglutathione hydrolase
• Tacrolimus Binding Proteins
• Tacrolimus

Topics

• Animals
• Autophagy (drug effects, physiology)
• Cell Cycle (drug effects, physiology)
• Cell Line
• Cell Line, Tumor
• Chickens
• HEK293 Cells
• HeLa Cells
• Humans
• Mechanistic Target of Rapamycin Complex 1 (metabolism)
• Phosphorylation (drug effects, physiology)
• Pyrazoles (pharmacology)
• Signal Transduction (drug effects, physiology)
• Tacrolimus (pharmacology)
• Tacrolimus Binding Proteins (metabolism)
• Thiolester Hydrolases (metabolism)
• Ubiquitination (drug effects, physiology)