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The mouse RANKL gene locus is defined by a broad pattern of histone H4 acetylation and regulated through distinct distal enhancers
Author(s) -
Martowicz Melissa L.,
Meyer Mark B.,
Pike J. Wesley
Publication year - 2011
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.23123
Subject(s) - acetylation , enhancer , histone h4 , histone , locus (genetics) , gene , biology , genetics , microbiology and biotechnology , gene expression
RANKL is a stromal cell‐derived tumor necrosis factor (TNF)‐like factor that plays a primary role in osteoclast formation and function. Recent studies suggest that 1,25(OH) 2 D 3 induces Rankl expression via vitamin D receptor (VDR) interaction at several enhancers located up to 76 kb upstream of the gene's transcriptional start site (TSS). In the current studies, we explored these interactions further using ChIP–chip and RNA analysis. We confirm VDR and RXR binding to the five enhancers described previously and identify two additional sites, one located within the Rankl coding region. We also show that RNA polymerase II is recruited to these enhancers, most likely through transcription factors TBP, TFIIB, and TAF II 250. Interestingly, the recruitment of these factors leads to the production of RNA transcripts, although their role at present is unknown. We also discovered that histone H4 acetylation (H4ac) marks many upstream Rankl enhancers under basal conditions and that H4ac is increased upon 1,25(OH) 2 D 3 treatment. Surprisingly, the hormone also induces C/EBPβ binding across the Rankl locus. C/EBPβ binding correlates directly with increased H4ac activity following 1,25(OH) 2 D 3 treatment. Finally, elevated H4ac is restricted to an extended region located between two potential insulator sites occupied by CTCF and Rad21. These data suggest a mechanism whereby 1,25(OH) 2 D 3 functions via the VDR and C/EBPβ to upregulate Rankl expression. J. Cell. Biochem. 112: 2030–2045, 2011. © 2011 Wiley‐Liss, Inc.