title: Core 2 ß1,6-N-acetylglucosaminyltransferase-I, crucial for P-selectin ligand expression is controlled by a distal enhancer regulated by STAT4 and T-bet in CD4+ T helper cells 1.
abstract: P-selectin ligands (P-ligs) support the recruitment of lymphocytes into inflamed tissues. Binding to P-selectin is mediated by oligosaccharide groups synthesized by means of several glycosyltransferases including core 2 ß1,6-N-acetylglucosaminyltransferase-I (C2GlcNAcT-I), encoded by the gene Gcnt1. Using Gcnt1(-/-) Th1 cells, we show that C2GlcNAcT-I is crucial for inflammatory T cell homing in vivo. To understand the molecular regulation of Gcnt1 in CD4(+) T helper cells, we performed ChIP-on-chip experiments across the Gcnt1 locus assessing the chromatin structure in P-lig-expressing versus non-expressing CD4(+) T cells. This identified a distal region about 20kb upstream of the promoter where the presence of a H3K27me3 mark correlated with Gcnt1 repression. This region possessed IL-12-dependent enhancer activity in reporter assays, in accordance with preferential IL-12-dependent induction of Gcnt1 in vitro. STAT4 and T-bet cooperated in control of the enhancer activity. Deficiency in either one resulted in drastically reduced Gcnt1 mRNA expression in differentiated Th1 cells. While both STAT4 and T-bet were bound to the enhancer early after activation only T-bet binding persisted throughout the expansion phase after TCR signal cessation. This suggests sequential action of STAT4 and T-bet at the enhancer. In summary, we show that Gcnt1 transcription and subsequent P-lig induction in Th1 cells is governed by binding of STAT4 and T-bet to a distal enhancer and further regulated by epigenetic marks such as H3K27me3.
authors: Deutsches Rheumaforschungszentrum, Experimentelle Rheumatologie, Berlin, Germany.
authors: Maibritt
authors: M
authors: Mardahl
authors: Deutsches Rheumaforschungszentrum, Experimentelle Rheumatologie, Berlin, Germany.
authors: Micha F
authors: MF
authors: Schröter
authors: Deutsches Rheumaforschungszentrum, Experimentelle Rheumatologie, Berlin, Germany.
authors: Dirk
authors: D
authors: Engelbert
authors: Deutsches Rheumaforschungszentrum, Experimentelle Rheumatologie, Berlin, Germany.
authors: Matthias
authors: M
authors: Pink
authors: Walter Brendel Zentrum für Experimentelle Medizin, Ludwig-Maximilians-Universität, Munich, Germany.
authors: Markus
authors: M
authors: Sperandio
authors: Deutsches Rheumaforschungszentrum, Experimentelle Rheumatologie, Berlin, Germany.
authors: Alf
authors: A
authors: Hamann
authors: Deutsches Rheumaforschungszentrum, Experimentelle Rheumatologie, Berlin, Germany; Charité Universitätsmedizin Berlin, Medizinische Klinik für Gastroenterologie, Infektiologie und Rheumatologie, Hindenburgdamm 30, 12200 Berlin, Germany. Electronic address: uta.syrbe@charite.de.
authors: Uta
authors: U
authors: Syrbe
mesh_terms: D000818:Animals; D002469:Cell Separation; D002634:Chemotaxis, Leukocyte / Q000276:immunology*; D047369:Chromatin Immunoprecipitation; D004742:Enhancer Elements, Genetic / Q000276:immunology; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000276:immunology*; D055786:Gene Knockout Techniques; D008213:Lymphocyte Activation / Q000276:immunology; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D017351:N-Acetylglucosaminyltransferases / Q000096:biosynthesis*; D016133:Polymerase Chain Reaction; D050797:STAT4 Transcription Factor / Q000276:immunology / Q000378:metabolism; D020825:T-Box Domain Proteins / Q000276:immunology / Q000378:metabolism; D018417:Th1 Cells / Q000276:immunology / Q000378:metabolism*; D000098815:T-bet Transcription Factor
chemical_list: D008562:Membrane Glycoproteins; C084882:P-selectin ligand protein; D050797:STAT4 Transcription Factor; C494091:Stat4 protein, mouse; D020825:T-Box Domain Proteins; D000098815:T-bet Transcription Factor; D017351:N-Acetylglucosaminyltransferases; C053024:beta-1,3-galactosyl-O-glycosyl-glycoprotein beta-1,6-acetylglucosaminyl transferase
keywords: Epigenetics; Gene regulation; Glycosyltransferase; Histone methylation; Homing; Lymphocyte