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Characterization of the human interleukin-5 gene promoter: involvement of octamer binding sites in the gene promoter activity

Abstract : To investigate the human interleukin (IL)-5 gene promoter, we have constructed a plasmid with the firefly luciferase reporter gene linked to human IL-5 5' flanking sequence (nucleotides -507 to +44). We have used this plasmid to transfect the mouse EL4 T cell line, which can, under certain conditions, produce IL-5 transcripts. Phorbol 12-myristate 13-acetate, A23187 and N6, 2'-O-dibutyryl-adenosine 3':5'-cyclic monophosphate co-stimulation of EL4 cells transfected with the human IL-5/luciferase reporter gene construct resulted in maximal induction of the luciferase gene. Deletion analysis of the IL-5 promoter revealed the presence of negative regulatory elements between nucleotides -404 and -312 and two regions, located between nucleotides -312 and -227 and between nucleotides -80 and -35, that are involved in the positive regulation of the IL-5 promoter. Using electrophoretic mobility shift assays, we show that the positive element located between nucleotides -312 and -227 involves the binding of factors antigenically related to Oct1, Oct2A and Oct2B, to a perfect octamer motif located at position -244/-237. Introduction of three point mutations in the octamer motif of the IL-5/luciferase reporter gene plasmid, which results in the loss of competition for the factors binding to the IL-5 promoter sequence, reduced the production of luciferase from stimulated, transfected EL4 cells, by 90%. Octamer factors can also bind within the second positive regulatory region.
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https://hal-univ-tours.archives-ouvertes.fr/hal-02414913
Contributor : Valérie Gouilleux <>
Submitted on : Monday, December 16, 2019 - 6:46:50 PM
Last modification on : Friday, January 10, 2020 - 4:12:10 PM

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Valérie Gruart-Gouilleux, Peter Engels, Michael Sullivan. Characterization of the human interleukin-5 gene promoter: involvement of octamer binding sites in the gene promoter activity. European Journal of Immunology, Wiley-VCH Verlag, 1995, 25 (5), pp.1431-1435. ⟨10.1002/eji.1830250544⟩. ⟨hal-02414913⟩

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