title: Multiple interferon regulatory factor and NF-κB sites cooperate in mediating cell-type- and maturation-specific activation of the human CD83 promoter in dendritic cells.
abstract: CD83 is one of the best-known surface markers for fully mature dendritic cells (mature 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, chromatin immunoprecipitation (ChIP)-on-chip microarray, biocomputational, reporter, electrophoretic mobility shift assay (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 have not been reported in this arrangement for any other gene so far. Notably, these DNA regions contain a complex framework of interferon regulatory factor (IRF)- and NF-κB transcription factor-binding sites mediating their arrangement. Mutation of any of the IRF-binding sites resulted in a significant loss of promoter activity, whereas overexpression of NF-κB 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 not only contributes to the knowledge of 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.
authors: Department of Dermatology, University Hospital Erlangen, Erlangen, Germany.
authors: Marcello F
authors: MF
authors: Stein
authors: Stefan
authors: S
authors: Lang
authors: Thomas H
authors: TH
authors: Winkler
authors: Andrea
authors: A
authors: Deinzer
authors: Sebastian
authors: S
authors: Erber
authors: Dirk M
authors: DM
authors: Nettelbeck
authors: Elisabeth
authors: E
authors: Naschberger
authors: Ramona
authors: R
authors: Jochmann
authors: Michael
authors: M
authors: Stürzl
authors: Robert K
authors: RK
authors: Slany
authors: Thomas
authors: T
authors: Werner
authors: Alexander
authors: A
authors: Steinkasserer
authors: Ilka
authors: I
authors: Knippertz
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D003713:Dendritic Cells / Q000378:metabolism*; D004742:Enhancer Elements, Genetic / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics* / Q000378:metabolism; D050835:Interferon Regulatory Factors / Q000235:genetics* / Q000378:metabolism; D007438:Introns; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D041681:NIH 3T3 Cells; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics*; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D007136:Immunoglobulins; D050835:Interferon Regulatory Factors; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D014157:Transcription Factors; D000097802:CD83 Antigen