title: HuR post-transcriptionally regulates TNF-α-induced IL-6 expression in human pulmonary microvascular endothelial cells mainly via tristetraprolin.
abstract: HuR and tristetraprolin (TTP) are both RNA-binding proteins, which are characterized as binding to the AU-rich elements (AREs) in the 3'-untranslated regions (3'-UTRs) of target mRNAs. Studies have shown that some ARE-containing mRNAs are stabilized by HuR, whereas are destabilized by TTP. Our previous study showed that HuR upregulated tumor necrosis factor-α (TNF-α)-induced interleukin-6 (IL-6) expression by stabilizing its mRNA in human pulmonary microvascular endothelial cells (HPMECs). Considering IL-6 mRNA has AREs, we decided to examine whether TTP was also involved in the regulation of TNF-α-induced IL-6 expression in HPMECs and whether HuR and TTP influenced each other at protein and mRNA level. Here, we report that TTP silencing increased IL-6 levels. HuR silencing increased TTP expression. TTP had no effect on HuR expression and subcellular localization. Compared to TTP silencing alone, double knockdown of HuR and TTP did not significantly reduce IL-6 release. The RNA-binding protein immunoprecipitation (RIP) results further showed that TTP but not HuR bound to intracellular IL-6 mRNA in HPMECs. We demonstrate for the first time that HuR post-transcriptionally regulates IL-6 expression mainly via TTP.
authors.affiliation: Department of Respiratory and Critical Care Medicine, Jinling Hospital, Nanjing University School of Medicine, 305 East Zhongshan Road, Nanjing 210002, China.
authors.forename: Jia-Xin
authors.initials: JX
authors.lastname: Shi
authors.forename: Xin
authors.initials: X
authors.lastname: Su
authors.forename: Jin
authors.initials: J
authors.lastname: Xu
authors.forename: Wei-Yun
authors.initials: WY
authors.lastname: Zhang
authors.forename: Yi
authors.initials: Y
authors.lastname: Shi
mesh_terms: D020413:3' Untranslated Regions; D002460:Cell Line; D051959:ELAV Proteins / Q000235:genetics / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism*; D020868:Gene Silencing; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis*; D008168:Lung / Q000098:blood supply / Q000378:metabolism; D055806:Microvessels / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D051816:Tristetraprolin / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D051959:ELAV Proteins; C508600:IL6 protein, human; D015850:Interleukin-6; D034741:RNA, Small Interfering; D051816:Tristetraprolin; D014409:Tumor Necrosis Factor-alpha; C497552:ZFP36 protein, human