Mass Spectrometry-based Identification of GP73 Interacting Proteins Reveals Its Regulatory Role on RNA Splicing Efficiency
10.13865/j.cnki.cjbmb.2025.02.1432
- VernacularTitle:基于相互作用蛋白质的质谱鉴定揭示高尔基体蛋白73对RNA剪接效率的调控作用
- Author:
Chang ZHANG
1
;
Mu-Yi LIU
;
Meng-Xin YANG
;
Lu-Ming WAN
;
Hui ZHONG
;
Cong-Wen WEI
Author Information
1. 安徽大学物质科学与信息技术研究院健康科学与技术研究所,合肥 230601
- Publication Type:Journal Article
- Keywords:
protein-protein interactions;
Golgi protein 73(GP73);
co-immunoprecipitation mass spec-trum;
bioinformatics analysis;
nuclear localization;
splicing efficiency
- From:
Chinese Journal of Biochemistry and Molecular Biology
2025;41(3):404-414
- CountryChina
- Language:Chinese
-
Abstract:
Protein-protein interactions play an extremely important role in the biochemical functions of cells,and in-depth analysis of protein interactions is the key to understanding cellular life activities.In this study,we systematically mined the interacting proteins of Golgi protein 73(GP73)using classical immunoprecipitation combined with mass spectrometry,and sought to further analyze the molecular func-tion of GP73.Hepatocellular carcinoma cell line HepG2 was selected,and a stable cell line overexpress-ing GP73-3Flag was constructed using lentiviral infection technology.A total of 78 high-confidence GP73 interacting proteins were identified by immunoprecipitation coupled with mass spectrometry.Bioinformat-ics analyses suggested that GP73 interacted with nearly 40 cytosolic proteins and participated in the bio-logical processes of RNA transport,splicing,and translation.Further immunofluorescence and cytosolic protein isolation experiments confirmed the cytosolic localization of GP73 in a variety of tumor cells.Based on the 78 interacting proteins,we further screened protein interaction networks related to mRNA splicing and verified the existence of interactions between GP73 and seven proteins,including HNRN-PH3,SMN1,RBM14,andNCBP1,by co-immunoprecipitation experiments.In addition,minigene spli-cing assay results indicated that GP73 inhibited the splicing efficiency of pre-mRNA by cells.This study contributes to the expansion of knowledge regarding the function of GP73 and aids in elucidating its criti-cal role in cell biology and its potential association with diseases.
