Methylation of an intragenic alternative promoter regulates transcription of GARP

Biochim Biophys Acta. 2016 Feb;1859(2):223-34. doi: 10.1016/j.bbagrm.2015.11.003. Epub 2015 Nov 14.

Abstract

Alternative promoter usage has been proposed as a mechanism regulating transcriptional and translational diversity in highly elaborated systems like the immune system in humans. Here, we report that transcription of human glycoprotein A repetitions predominant (GARP) in regulatory CD4 T cells (Tregs) is tightly regulated by two alternative promoters. An intragenic promoter contains several CpGs and acts as a weak promoter that is demethylated and initiates transcription Treg-specifically. The strong up-stream promoter containing a CpG-island is, in contrast, fully demethylated throughout tissues. Transcriptional activity of the strong promoter was surprisingly down-regulated upon demethylation of the weak promoter. This demethylation-induced transcriptional attenuation regulated the magnitude of GARP expression and correlated with disease activity in rheumatoid arthritis. Treg-specific GARP transcription was initiated by synergistic interaction of forkhead box protein 3 (Foxp3) with nuclear factor of activated T cells (NFAT) and was underpinned by permissive chromatin remodeling caused by release of the H3K4 demethylase, PLU-1. Our findings describe a novel function of alternative promoters in regulating the extent of transcription. Moreover, since GARP functions as a transporter of transforming growth factor β (TGFβ), a cytokine with broad pleiotropic traits, GARP transcriptional attenuation by alternative promoters might provide a mechanism regulating peripheral TGFβ to avoid unwanted harmful effects.

Keywords: Alternative promoter; DNA methylation; GARP; Gene transcription; Transcriptional interference; Treg.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin Assembly and Disassembly / genetics
  • CpG Islands
  • DNA Methylation / genetics*
  • Forkhead Transcription Factors / genetics*
  • Gene Expression Regulation
  • Humans
  • Jumonji Domain-Containing Histone Demethylases / genetics
  • Jurkat Cells
  • Membrane Proteins / biosynthesis
  • Membrane Proteins / genetics*
  • NFATC Transcription Factors / genetics*
  • Nuclear Proteins / genetics
  • Promoter Regions, Genetic*
  • Repressor Proteins / genetics
  • T-Lymphocytes, Regulatory / metabolism
  • Transcription, Genetic*

Substances

  • FOXP3 protein, human
  • Forkhead Transcription Factors
  • LRRC32 protein, human
  • Membrane Proteins
  • NFATC Transcription Factors
  • Nuclear Proteins
  • Repressor Proteins
  • Jumonji Domain-Containing Histone Demethylases
  • KDM5B protein, human