Posttranscriptional regulation of IL-13 in T cells: Role of the RNA-binding protein HuR

Vincenzo Casolaro, Xi Fang, Brian Tancowny, Jinshui Fan, Fan Wu, Subramanya Srikantan, S. Yukiko Asaki, Umberto De Fanis, Shau Ku Huang, Myriam Gorospe, Ulus X. Atasoy, Cristiana Stellato

Research output: Contribution to journalArticlepeer-review

44 Scopus citations


Background: IL-13, a critical cytokine in allergy, is regulated by as-yet-elusive mechanisms. Objective: We investigated IL-13 posttranscriptional regulation by HuR, a protein associating with adenylate-uridylate-rich elements in the 3′ untranslated regions (UTRs) of mRNA, promoting mRNA stability and translation. Methods: IL-13 mRNA decay was monitored in human TH2-skewed cells by using the transcriptional inhibitor actinomycin D. The IL-13 3′UTR was subcloned into an inducible β-globin reporter transiently expressed in H2 cells in the absence or presence of overexpressed HuR. Association of HuR with IL-13 mRNA was detected by means of immunoprecipitation of ribonucleoprotein complexes and a biotin pull-down assay. The effects of HuR transient overexpression and silencing on IL-13 expression were investigated. Results: IL-13 mRNA half-life increased significantly in restimulated TH2-skewed cells compared with baseline values. Decay of β-globin mRNA was significantly faster in H2 cells transfected with the IL-13 3′UTR-containing plasmid than in those carrying a control vector. HuR overexpression increased the β-globin IL-13 3′UTR reporter half-life. Significant enrichment of IL-13 mRNA was produced by means of immunoprecipitation of Jurkat cell ribonucleoprotein complexes with anti-HuR. HuR binding to the IL-13 3′UTR was confirmed by means of pull-down assay of biotin-labeled RNA probes spanning the IL-13 3′UTR. Two-dimensional Western blot analysis showed stimulus-induced posttranslational modification of HuR. In Jurkat cells mitogen-induced IL-13 mRNA was significantly affected by HuR overexpression and silencing. Conclusions: Mitogen-induced IL-13 expression involves changes in transcript turnover and a change in phosphorylation of HuR and its association with the mRNA 3′UTR.

Original languageEnglish (US)
Pages (from-to)853-859.e4
JournalJournal of Allergy and Clinical Immunology
Issue number4
StatePublished - Apr 2008


  • Asthma
  • IL-13
  • RNA-binding proteins
  • T cells
  • T2 cytokines
  • inflammation
  • mRNA turnover
  • posttranscriptional gene regulation

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology


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