Long-Range Enhancer Differentially Regulated by c-Jun and JunD Controls Peptidylarginine Deiminase-3 Gene in Keratinocytes

UMR 5165, CNRS-Toulouse III University, CHU Purpan, Place du Dr Baylac TSA4003, 31059 Toulouse cedex 9, France.
Journal of Molecular Biology (Impact Factor: 4.33). 11/2008; 384(5):1048-57. DOI: 10.1016/j.jmb.2008.10.019
Source: PubMed


Long-range cis elements are critical regulators of transcription, particularly for clustered paralogous genes. Such are the five PADI genes in 1p35-36 encoding peptidylarginine deiminases, which catalyze deimination, a Ca2+-dependent post-translational modification. Deimination has been implicated in the pathophysiology of severe human diseases such as multiple sclerosis and rheumatoid arthritis. The PADI genes present different expression patterns. PADI1-3 are expressed in the epidermis, with increased expression levels in the most differentiated keratinocytes. Previous studies on PADI proximal promoters failed to explain such specificity of expression. We identified a conserved intergenic sequence in the PADI locus (IG1), which may play a role in PADI transcriptional regulation. In this work, we identified two DNase I.hypersensitive sites located in IG1, PAD intergenic enhancer segment 1 (PIE-S1) and PIE-S2, which act in synergy as a bipartite enhancer of the PADI3 and probably PADI1 promoters in normal human epidermal keratinocytes differentiated by a high-calcium-containing medium (1.5 mM). PIE-S1 and PIE-S2 present all the hallmarks of transcriptional enhancers: orientation-independence, copy-number dependence and cell-type specificity. PIE-S1 and PIE-S2 comprise conserved putative binding sites for MIBP1/RFX1 and activator protein 1, respectively. Deletion mutant screening revealed that these sites are crucial for the enhancer activity. Furthermore, chromatin immunoprecipitation assays evidenced differential binding of JunD or c-Jun on the activator protein 1 site depending on the cell differentiation state. Our results reveal the molecular bases of the expression specificity of PADI1 and PADI3 during keratinocyte differentiation through a long-range enhancer and support a model of PADI gene regulation depending on c-Jun-JunD competition.

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    • "The spatially complex expression patterns displayed by many genes often rely on the presence of remote regulatory elements that modulate the activity of gene promoters in a tissue-specific or stimulus-inducible manner [47]. In addition, there is evidence that the interaction of these remote regulatory elements with the promoters of the genes they regulate is highly specific [48,49,50]. In the past, the identification of these remote regulatory elements by deletion analysis was very labour intensive and time consuming. "
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