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Identification of an histone H3 acetylated/K4-methylated-bound intragenic enhancer regulatory for urokinase receptor expression

The transcriptionally regulated urokinase-type plasminogen activator receptor (u-PAR) contributes to cancer progression. Although previous studies have identified multiple 5′ regulatory elements, these cis motifs cannot fully account for u-PAR expression prompting a search for hitherto uncharacteriz...

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Main Authors: Wang, H. (Author) , Yan, C. (Author) , Asangani, Irfan A. (Author) , Allgayer, Heike (Author) , Boyd, D. D. (Author)
Format: Article (Journal)
Language:English
Published: 2007
In: Oncogene
Year: 2007, Volume: 26, Issue: 14, Pages: 2058-2070
ISSN:1476-5594
DOI:10.1038/sj.onc.1210003
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1038/sj.onc.1210003
Verlag, lizenzpflichtig, Volltext: https://www.nature.com/articles/1210003
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Author Notes:H. Wang, C. Yan, I. Asangani, H. Allgayer and D.D. Boyd

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520 |a The transcriptionally regulated urokinase-type plasminogen activator receptor (u-PAR) contributes to cancer progression. Although previous studies have identified multiple 5′ regulatory elements, these cis motifs cannot fully account for u-PAR expression prompting a search for hitherto uncharacterized regulatory elements. DNase I hypersensitivity and chromatin immunoprecipitation assays using u-PAR-expressing colon cancer cells indicated a hypersensitive region (+665/+2068) in intron 1 enriched with acetylated histone 3 (H3) and H3 methylated at lysine 4, markers of regulatory regions. The +665/+2068 region increased transcription from a u-PAR-promoter in an orientation- and distance-independent manner fulfilling the criteria of an enhancer. Optimal stimulation of the u-PAR promoter by phorbol ester required this enhancer. Systematic truncations combined with DNase I footprinting revealed two protected regions (+1060/+1099 and +1123/+1134) with deletion of the latter practically abolishing enhancer activity. The +1123/+1134 region harbored non-consensus activator protein-1 and Ets1 binding sites bound with c-Jun (and/or the related JunD/JunB) and c-Fos (and/or the related FosB/Fra-1/Fra-2) as revealed with chromatin immunoprecipitation. Further, nuclear extract from resected colon cancers showed elevated protein binding to a +1123/+1134-spanning probe coordinate with elevated u-PAR protein. Thus, we have defined a novel intragenic enhancer in the u-PAR gene required for constitutive and inducible expression. 
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