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Multiple IRF- and NFkB-sites cooperate in mediating cell type- and maturation-specific activation of the human CD83 promoter in dendritic cells.


Stein MF, Lang S, Winkler TH, Deinzer A, Erber S, Nettelbeck DM, Naschberger E, Jochmann R, Stürzl M, Slany RK, Werner T, Steinkasserer A, Knippertz I

CD83 is one of the best-known surface marker for fully mature dendritic cells (DCs) and its cell type- and maturation-specific regulation makes the CD83 promoter an interesting tool for the genetic modulation of DCs. To determine the mechanisms regulating this DC- and maturation-specific CD83 expression, ChIP-on-chip-microarray-, biocomputational-, reporter-, EMSA- and ChIP-analyses were performed. These studies led to the identification of a ternary transcriptional activation complex composed of an upstream regulatory element, a minimal promoter and an enhancer which in this arrangement have not been reported for any other gene so far. Notably, these DNA regions contain a complex framework of IRF- and NFkB-transcription factor binding sites mediating their arrangement. Mutation of any of the IRF-binding sites resulted in a significant loss of promoter activity, whereas over-expression of NFkB-transcription factors clearly enhanced transcription. We identified IRF-1, IRF-2, IRF-5, p50, p65 and cRel to be involved in regulating maturation-specific CD83 expression in DCs. Therefore, the characterization of this promoter complex contributes not only to the knowledge on DC-specific gene regulation, but also suggests the involvement of a transcriptional module with binding sites separated into distinct regions in transcriptional activation as well as cell type- and maturation-specific transcriptional targeting of DCs.

Tags
Bioruptor
Chromatin Shearing
ChIP-qPCR

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Published
April, 2013

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