Proteomic profiling of eIF3a conditional knockout mice

Wei Zhuo; Juan Chen; Shilong Jiang; Juyan Zheng; Hanxue Huang; Pan Xie; Wei Li; Mengrong Lei; Jiye Yin; Ying Gao; Zhaoqian Liu

doi:10.3389/fmolb.2023.1160063

Proteomic profiling of eIF3a conditional knockout mice

Front Mol Biosci. 2023 Apr 19:10:1160063. doi: 10.3389/fmolb.2023.1160063. eCollection 2023.

Authors

Wei Zhuo^{1

2}, Juan Chen³, Shilong Jiang³, Juyan Zheng^{1

2}, Hanxue Huang^{1

2}, Pan Xie^{1

2}, Wei Li^{1

2}, Mengrong Lei^{1

2}, Jiye Yin^{1

2}, Ying Gao^{1

4}, Zhaoqian Liu^{1

2}

Affiliations

¹ Department of Clinical Pharmacology, Hunan Key Laboratory of Pharmacogenetics, National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China.
² Engineering Research Center for Applied Technology of Pharmacogenomics of Ministry of Education, Institute of Clinical Pharmacology, Central South University, Changsha, China.
³ Departments of Pharmacy, Xiangya Hospital, Central South University, Changsha, China.
⁴ Departments of Gerontology, Xiangya Hospital, Central South University, Changsha, China.

Abstract

Eukaryotic translation initiation factor 3 subunit A (eIF3a) is the largest subunit of the eukaryotic translation initiation factor 3 (eIF3). eIF3a plays an integral role in protein biosynthesis, hence impacting the onset, development, and treatment of tumors. The proteins regulated by eIF3a are still being explored in vivo. In this study, a Cre-loxP system was used to generate eIF3a conditional knockout mice. Tandem mass tag (TMT) labeling with LC-MS/MS analysis was used to identify differentially expressed proteins (DEPs) in fat, lungs, skin, and spleen tissue of the eIF3a knockout mice and controls. Bioinformatics analysis was then used to explore the functions and molecular signaling pathways of these protein landscapes. It was observed that eIF3a is essential for life sustenance. Abnormal tissue pathology was found in the lungs, fat, skin, spleen, and thymus. In total, 588, 210, 324, and 944 DEPs were quantified in the lungs, fat, skin, and spleen, respectively, of the eIF3a knockout mice as compared to the control. The quantified differentially expressed proteins were tissue-specific, except for eight proteins shared by the four tissues. A broad range of functions for eIF3a, including cellular signaling pathway, immune response, metabolism, defense response, phagocytes, and DNA replication, has been revealed using bioinformatics analysis. Herein, several pathways related to oxidative stress in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, including nitrogen metabolism, peroxisome, cytochrome P450 drug metabolism, pyruvate metabolism, PPAR signaling pathway, phospholipase D signaling pathway, B-cell receptor signaling pathway, ferroptosis, and focal adhesion, have been identified. Collectively, this study shows that eIF3a is an essential gene for sustaining life, and its downstream proteins are involved in diverse novel functions beyond mRNA translational regulation.

Keywords: eIF3a; knockout mice; oxidative stress; protein landscape; proteomics.

Grants and funding

This work was supported by the National Natural Science Foundation of China (81874327 and 82173901), the Project Program of National Clinical Research Center for Geriatric Disorders (Xiangya Hospital, 2020LNJJ02) of China, the Science and Technology Program of Changsha (kh2003010) of China, and the Fundamental Research Funds for the Central Universities of Central South University (2017zzts225).