IL-10 inhibits transcription elongation of the human TNF gene in primary macrophages

Tim Smallie, Giuseppe Ricchetti, Nicole J Horwood, Marc Feldmann, Andrew R Clark, Lynn M Williams

Research output: Contribution to journalArticlepeer-review

76 Citations (Scopus)

Abstract

IL-10 plays a central nonredundant role in limiting inflammation in vivo. However, the mechanisms involved remain to be resolved. Using primary human macrophages, we found that IL-10 inhibits selected inflammatory genes, primarily at a level of transcription. At the TNF gene, this occurs not through an inhibition of RNA polymerase II (Pol II) recruitment and transcription initiation but through a mechanism targeting the stimulation of transcription elongation by cyclin-dependent kinase (CDK) 9. We demonstrated an unanticipated requirement for a region downstream of the TNF 3' untranslated region (UTR) that contains the nuclear factor κB (NF-κB) binding motif (κB4) both for induction of transcription by lipopolysaccharide (LPS) and its inhibition by IL-10. IL-10 not only inhibits the recruitment of RelA to regions containing κB sites at the TNF gene but also to those found at other LPS-induced genes. We show that although IL-10 elicits a general block in RelA recruitment to its genomic targets, the gene-specific nature of IL-10's actions are defined through the differential recruitment of CDK9 and the control of transcription elongation. At TNF, but not NFKBIA, the consequence of RelA recruitment inhibition is a loss of CDK9 recruitment, preventing the stimulation of transcription elongation.

Original languageEnglish
Pages (from-to)2081-2088
Number of pages8
JournalJournal of Experimental Medicine
Volume207
Issue number10
Early online date30 Aug 2010
DOIs
Publication statusPublished - 27 Sep 2010

Keywords

  • 3' Untranslated Regions/genetics
  • Binding Sites/genetics
  • Cells, Cultured
  • Cyclin-Dependent Kinase 9/genetics
  • Humans
  • I-kappa B Proteins/genetics
  • Interleukin-10/immunology
  • Lipopolysaccharides/pharmacology
  • Macrophages/immunology
  • NF-KappaB Inhibitor alpha
  • NF-kappa B/genetics
  • Promoter Regions, Genetic
  • RNA Polymerase II/genetics
  • Transcription Factor RelA/genetics
  • Transcription, Genetic/drug effects
  • Transcriptional Activation
  • Tumor Necrosis Factor-alpha/genetics

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