Large-Scale Proteomic Identification of Targets of Cellular miR-197 Downregulated by Enterovirus A71

Wen-Fang Tang, Ru Ting Huang, Kun Yi Chien, Petrus Tang, Jim Tong Horng*

*此作品的通信作者

研究成果: 期刊稿件文章同行評審

4 引文 斯高帕斯(Scopus)

摘要

MicroRNAs are noncoding RNA species comprising 18-23 nucleotides that regulate host-virus interaction networks. Here, we show that enterovirus A71 infection in human rhabdomyosarcoma (RD) is regulated by miR-197 expression. Transfection of miR-197 mimic into RD cells inhibited virus replication by interfering with the viral RNA synthesis. We employed a combination of mass-spectrometry-based quantitative proteomics with the stable isotope labeling with amino acids in cell culture (SILAC) approach for the identification of the miR-197 target genes in RD cells and to investigate the differential expression of the prospective target proteins. A total of 1822 proteins were repeatedly identified in miR-197-transfected RD cells, 106 of which were predicted to have seed sites by TargetScan. Notably, seven of eight selected genes potentially related to viral replication and immune response were validated as direct miR-197 targets, using a luciferase 3′-untranslated region (UTR) reporter assay. The expression levels of three selected endogenous molecules (ITGAV, ETF1, and MAP2K1/MEK1) were significantly reduced when RD cells were transfected with a miR-197 mimic. Our results provide a comprehensive database of miR-197 targets, which might provide better insights into the understanding of host-virus interaction.

原文英語
頁(從 - 到)449-460
頁數12
期刊Journal of Proteome Research
18
發行號1
DOIs
出版狀態已出版 - 04 01 2019

文獻附註

Publisher Copyright:
© Copyright 2018 American Chemical Society.

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