Study on Regulation of mTORC1 Integrity and Activity by TRiC/CCT Complex
10.3870/j.issn.1672-0741.24.05.031
- VernacularTitle:TRiC/CCT复合体对mTORC1完整性及其活性的调控作用研究
- Author:
Xianhang YAN
1
;
Zhengtao QIAN
;
Chunfang YAO
Author Information
1. 江苏大学医学院,镇江 212013
- Publication Type:Journal Article
- Keywords:
mTORC1;
TRiC/CCT complex;
protein folding;
molecular chaperone
- From:
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
2025;54(1):73-79
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the possibility of mutual regulation between TRiC/CCT complex(TCP-1 ring complex/chaperonin containing TCP-1)and mTOR complex 1(mechanistic target of rapamycin complex 1).Methods co-immunoprecipi-tation and Western blot were adopted to explore the interaction between TRiC/CCT complex and mTORC1.Rapamycin was used to inhibit the activity of mTORC 1.Soluble and insoluble cell protein samples were prepared,and the changes in protein ex-pression levels of TRiC/CCT complex-assisted folding were compared.The changes were observed via non-denaturing gel elec-trophoresis to explore the influences of mTORC1 on the integrity of TRiC/CCT complex.The CCT2 or CCT5 gene in HEK-293T cells was silenced by lentivirus-mediated shRNA to explore the influences of TRiC/CCT complex on the expression of key components of mTORC1 and the phosphorylation level of downstream substrates.Results Interaction between each subunit of TRiC/CCT complex and mTORC1 was proved by co-immunoprecipitation experiment.The activity of mTORC1 was inhibited by rapamycin.Expression levels of β-actin,β-Tubulin,and STAT3 insoluble proteins were not significantly increased,and the molecular size of the complex did not change significantly in the non-denaturing electrophoresis gel,indicating that mTORC1 had no regulative effect on formation of TRiC/CCT complex.CCT2 or CCT5 was silenced in HEK-293T.The protein expres-sion levels of mTOR,Raptor,and mLST8 were decreased,and the phosphorylation levels of p70S6K and Akt were decreased,re-vealing that the TRiC/CCT complex had a regulative effect on mTORC1 and downstream molecules.Conclusion The protein expression level of each component of mTORC1 was regulated by TRiC/CCT complex,thus affecting the phosphorylation level and activity status of the downstream substrates.
