miR-365, a novel negative regulator of interleukin-6 gene expression, is cooperatively regulated by Sp1 and NF-kappaB

J Biol Chem. 2011 Jun 17;286(24):21401-12. doi: 10.1074/jbc.M110.198630. Epub 2011 Apr 25.

Abstract

Interleukin-6 (IL-6) is a pleiotropic cytokine that plays a central role in host defense. IL-6 expression can be regulated at both a transcriptional and a post-transcriptional level. We used a combination of bioinformatics and experimental techniques to demonstrate that the miR-365 is a direct negative regulator of IL-6. Overexpression of miR-365 mimics decreased activity of a luciferase reporter containing the IL-6 3'-UTR and led to repression of IL-6 protein. In contrast, ectopic expression of a miR-365 inhibitor elevated IL-6 expression. The negative regulation of miR-365 was strictly dependent on a microRNA binding element in the 3'-UTR of IL-6 mRNA. Deletion mutant analysis of the miR-365 promoter showed that two transcription factors, Sp1 and NF-κB, are essential for the transcriptional regulation of miR-365. We also demonstrate that the MAPK/ERK pathway contributes to the regulation of miR-365. Furthermore, miR-365 exhibited a greater negative regulatory effect on IL-6 than hsa-let-7a, a previously identified microRNA negatively regulating IL-6. Taken together, our results show that miR-365 is a novel negative regulator of IL-6.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Base Sequence
  • Gene Expression Regulation*
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Interleukin-6 / biosynthesis*
  • MicroRNAs / biosynthesis*
  • MicroRNAs / physiology*
  • Molecular Sequence Data
  • NF-kappa B / metabolism*
  • Plasmids / metabolism
  • Protein Binding
  • RNA, Small Interfering / metabolism
  • Sequence Homology, Nucleic Acid
  • Sp1 Transcription Factor / metabolism*

Substances

  • 3' Untranslated Regions
  • Interleukin-6
  • MIRN365 microRNA, human
  • MicroRNAs
  • NF-kappa B
  • RNA, Small Interfering
  • Sp1 Transcription Factor