Proteomics

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A large-scale proteomic identification of targets of cellular miR-197 downregulated by enterovirus A71


ABSTRACT: MicroRNAs are noncoding RNA species comprising 18–23 nucleotides that regulate host-virus interaction networks. Here, we showed that enterovirus A71 infection in human rhabdomyosarcoma (RD) involved miR-197 expression. miR-197 can regulate virus replication in the context of viral RNA synthesis via the transfection of its mimic into RD cells. We employed a mass spectrometry-based quantitative proteomic stable isotope labeling with amino acids in cell culture (SILAC) approach for the identification of miR-197 target genes by transfecting mimetic miR-197 into RD cells, and the differential expression of prospective target proteins was identified. A total of 1,822 genes were repeatedly and considerably downregulated in miR-197-transfected RD cells, 106 of which were predicted to have seed sites by TargetScan. Seven of the selected 8 genes potentially related to viral replication and immune response were confidently validated as direct miR-197 targets using a luciferase (untranslated region (3'-UTR)) reporter assay. The expression of three selected endogenous molecules (ITGAV, ETF1, and MAP2K1 (MEK1)) was significantly reduced when RD cells were transfected with an miR-197 mimic. Our results provide a database of miR-197 targets, which is potentially of interest in the area of viral pathogenesis and other research fields.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

DISEASE(S): Rhabdomyosarcoma

SUBMITTER: WEN-FANG TANG  

LAB HEAD: Jim-Tong Horng

PROVIDER: PXD010585 | Pride | 2018-10-24

REPOSITORIES: Pride

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Publications

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

Tang Wen-Fang WF   Huang Ru-Ting RT   Chien Kun-Yi KY   Tang Petrus P   Horng Jim-Tong JT  

Journal of proteome research 20181029 1


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  ...[more]

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