Characterization of the human ephrin-A4 promoter.
Expression of the ephrin-A4 ligand, a family member of ligands binding the Eph receptor tyrosine kinases, is induced after an antigen-receptor stimulation of lymphocytes. To understand the transcription regulation of the ephrin-A4 gene, its promoter was identified and regulating elements were characterized. The ephrin-A4 promoter contains cis elements directing the cell-specific expression. By deletion studies, three specific regions, which were contributing to the transcription activity in lymphoid cells, were localized. In one of these regions, an inverted CCAAT box was identified and shown to bind the transcription activator nuclear factor-Y (NF-Y). The importance of NF-Y binding for the ephrin-A4 promoter activity is shown by a total abrogation of promoter activity after destruction of its binding site. NF-Y binding and activity are also crucially dependent on the integrity of the surrounding sequence. In addition, electrophoretic mobility-shift assay and serial-mutation analysis of the two remaining regulating regions revealed cis regulatory elements contributing to the transcription activity of the ephrin-A4 promoter.
Munthe
Else
E
Department of Immunology, The Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway. elsem@labmed.uio.no
Aasheim
Hans-Christian
HC
D020121:5' Untranslated Regions / Q000235:genetics; D001402:B-Lymphocytes; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D017931:DNA Primers; D036385:Ephrin-A4; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic
D020121:5' Untranslated Regions; D017931:DNA Primers; D036385:Ephrin-A4; D008565:Membrane Proteins; D008156:Luciferases