Proteomic screening identifies RPLp2 as a specific regulator for the translation of coronavirus

• Published in: International journal of biological macromolecules 2023-03, Vol.230, p.123191-123191, Article 123191
• Main Authors: Dong, Hui-Jun, Wang, Jing, Zhang, Xiu-Zhong, Li, Cui-Cui, Liu, Jian-Feng, Wang, Xiao-Jia
• Format: Article
• Language: English
• Subjects: Coronavirus; Coronavirus - genetics; Coronavirus Infections; Eukaryotic Initiation Factor-4E - metabolism; Eukaryotic translation initiation factor eIF4E; Humans; PEDV; Protein Biosynthesis; Proteomics; Ribosomal protein RPLp2; RNA, Messenger - genetics; RNA, Messenger - metabolism; Translation initiation; Viral nucleocapsid protein
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2023.123191

Viral mRNA of coronavirus translates in an eIF4E-dependent manner, and the phosphorylation of eIF4E can modulate this process, but the role of p-eIF4E in coronavirus infection is not yet entirely evident. p-eIF4E favors the translation of selected mRNAs, specifically the mRNAs that encode proteins associated with cell proliferation, inflammation, the extracellular matrix, and tumor formation and metastasis. In the present work, two rounds of TMT relative quantitative proteomics were used to screen 77 cellular factors that are upregulated upon infection by coronavirus PEDV and are potentially susceptible to a high level of p-eIF4E. PEDV infection increased the translation level of ribosomal protein lateral stalk subunit RPLp2 (but not subunit RPLp0/1) in a p-eIF4E-dependent manner. The bicistronic dual-reporter assay and polysome profile showed that RPLp2 is essential for translating the viral mRNA of PEDV. RNA binding protein and immunoprecipitation assay showed that RPLp2 interacted with PEDV 5′UTR via association with eIF4E. Moreover, the cap pull-down assay showed that the viral nucleocapsid protein is recruited in m7GTP-precipitated complexes with the assistance of RPLp2. The heterogeneous ribosomes, which are different in composition, regulate the selective translation of specific mRNAs. Our study proves that viral mRNA and protein utilize translation factors and heterogeneous ribosomes for preferential translation initiation. This previously uncharacterized process may be involved in the selective translation of coronavirus.