title: Transcriptional regulation of the human TNFSF11 gene in T cells via a cell type-selective set of distal enhancers.
abstract: In addition to osteoblast lineage cells, the TNF-like factor receptor activator of NF-κB ligand (RANKL) is expressed in both B and T cells and may play a role in bone resorption. Rankl gene (Tnfsf11) expression in mouse T cells is mediated through multiple distal elements marked by increased transcription factor occupancy, histone tail acetylation, and RNA polymerase II recruitment. Little is known, however, of the regulation of human TNFSF11 in T cells. Accordingly, we examined the consequence of T cell activation on the expression of this factor both in Jurkat cells and in primary human T cells. We then explored the mechanism of this regulation by scanning over 400 kb of DNA surrounding the TNFSF11 locus for regulatory enhancers using ChIP-chip analysis. Histone H3/H4 acetylation enrichment identified putative regulatory regions located between -170 and -220 kb upstream of the human TNFSF11 TSS that we designated the human T cell control region (hTCCR). This region showed high sequence conservation with the mouse TCCR. Inhibition of MEK1/2 by U0126 resulted in decreased RANKL expression suggesting that stimulation through MEK1/2 was a prerequisite. ChIP-chip analysis also revealed that c-FOS was recruited to the hTCCR as well. Importantly, both the human TNFSF11 D5a/b (RLD5a/b) enhancer and segments of the hTCCR mediated robust inducible reporter activity following TCR activation. Finally, SNPs implicated in diseases characterized by dysregulated BMD co-localized to the hTCCR region. We conclude that the hTCCR region contains a cell-selective set of enhancers that plays an integral role in the transcriptional regulation of the TNFSF11 gene in human T cells.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706.
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Bishop
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Wang
authors.forename: Heidi M
authors.initials: HM
authors.lastname: Coy
authors.forename: Mark B
authors.initials: MB
authors.lastname: Meyer
authors.forename: Jenny E
authors.initials: JE
authors.lastname: Gumperz
authors.forename: J Wesley
authors.initials: JW
authors.lastname: Pike
mesh_terms: D015519:Bone Density / Q000235:genetics; D002478:Cells, Cultured; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D006801:Humans; D019169:Jurkat Cells; D008213:Lymphocyte Activation; D048369:MAP Kinase Kinase 1 / Q000378:metabolism; D048370:MAP Kinase Kinase 2 / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide; D053245:RANK Ligand / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000378:metabolism*
chemical_list: D053245:RANK Ligand; C505688:TNFSF11 protein, human; C482128:MAP2K2 protein, human; D048369:MAP Kinase Kinase 1; D048370:MAP Kinase Kinase 2; C482125:MAP2K1 protein, human
keywords: C-FOS; ChIP-CHIP ANALYSIS; DISTAL ENHANCERS; EPIGENETIC MARKS; T CELL RANKL