Recruitment of Histone Methyltransferase Ehmt1 to Foxp3 TSDR Counteracts Differentiation of Induced Regulatory T Cells. Issue 19 (6th September 2019)

• Record Type: Journal Article
• Authors: Karl, Martin; Sommer, Christian; Gabriel, Christian H.; Hecklau, Katharina; Venzke, Melanie; Hennig, Anna Floriane; Radbruch, Andreas; Selbach, Matthias; Baumgrass, Ria
• Is Part Of: Journal of molecular biology. Volume 431:Issue 19(2019)
• Page Start: 3606
• Page End: 3625
• Publication Date: 2019-09-06
• Subjects: Cux1 cut like homeobox 1 -- CNS2 conserved non-coding sequence 2 -- Ehmt euchromatic histone-lysine N-methyltransferase -- PD-MS DNA pull-down with subsequent mass spectrometric analysis -- PD-WB DNA pull-down with subsequent Western Blot analysis -- Tcon conventional T cell -- (i/n)Treg (in vitro induced/natural) regulatory T cell -- TSDR Treg-specific demethylated region -- vitC vitamin C -- Wiz widely interspaced zinc finger motifs; epigenetics -- vitamin C -- T helper cell -- DNA-pull down -- mass spectrometry -- ascorbate; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2019.07.031
• Languages: English
• ISSNs: 0022-2836

Abstract

Abstract: Differentiation toward CD4 + regulatory T (Treg) cells is essentially dependent on an epigenetic program at Treg signature genes, which involves remodeling of the Treg-specific demethylated regions (TSDRs). In particular, the epigenetic status of the conserved non-coding sequence 2 of Foxp3 (Foxp3 TSDR) determines expression stability of the master transcription factor and thus Treg lineage identity. However, the molecular mechanisms controlling the epigenetic remodeling at TSDRs in Treg and conventional T cells are largely unknown. Using a combined approach of DNA pull-down and mass spectrometric analysis, we report a novel regulatory mechanism in which transcription factor Wiz recruits the histone methyltransferase Ehmt1 to Foxp3 TSDR. We show that both Wiz and Ehmt1 are crucial for shaping the region with the repressive histone modification H3K9me2 in conventional T cells. Consistently, knocking out either Ehmt1 or Wiz by CRISPR/Cas resulted in the loss of H3K9me2 and enhanced Foxp3 expression during iTreg differentiation. Moreover, the essential role of the Wiz–Ehmt1 interaction as observed at several TSDRs indicates a global function of Ehmt1 in the Treg differentiation program. Graphical Abstract: Unlabelled Image Highlights: Epigenetic remodeling of Foxp3 TSDR is known to be key for stable Treg cell phenotype. Ehmt1 maintains repressive histone mark H3K9me2 at Foxp3 TSDR in non-Treg cells. Recruitment of Ehmt1 is mediated by transcription factor Wiz. Knockout of Ehmt1 or Wiz by CRISPR/Cas promotes Treg differentiation. Ehmt1 activity at TSDRs is a basal control mechanism of Treg differentiation.