title: The activity of the human interleukin-5 conserved lymphokine element 0 is regulated by octamer factors in human cells.
abstract: Interleukin-5 (IL-5) controls the development of eosinophilia and contributes to a number of disease states including asthma. Expression of IL-5 is inducible under tight transcriptional regulation. This requires the contribution of several promoter elements; however, the conserved lymphokine element 0 (CLE0) in particular, is essential for expression of IL-5. In this study, we report the nuclear factors which regulate human IL-5 CLE0 activity in the human cell line PER-117. Using specific antibodies, we identified the transcriptional factors Oct-1 and Oct-2 binding to the 5' region of the CLE0 element. The involvement of Oct factors with CLE0 has not been reported previously in any of the lymphokines. In addition, the CLE0 element also appeared to complex with the transcriptional activator AP-1, consisting of the family members Jun D and Fra-2. We observed the binding of Oct-1 to be constitutive in comparison to Oct-2 and AP-1, both of which were induced in response to cell activation by PMA/A23187. Although the interaction of all three factors with CLE0 was closely linked and overlapping, residues critical to their binding were identified. We demonstrate, using site-directed mutagenesis and cotransfection experiments, that the CLE0 element is indispensable for IL-5 promoter activity and that Octamer factors contribute to the positive regulation of the hIL-5 gene.
authors.affiliation: TVWT Institute for Child Health Research, West Perth, Australia.
authors.forename: M A
authors.initials: MA
authors.lastname: Thomas
authors.forename: V A
authors.initials: VA
authors.lastname: Mordvinov
authors.forename: C J
authors.initials: CJ
authors.lastname: Sanderson
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D051863:Host Cell Factor C1; D006801:Humans; D015848:Interleukin-5 / Q000096:biosynthesis / Q000235:genetics*; D008222:Lymphokines / Q000096:biosynthesis / Q000235:genetics*; D050811:Octamer Transcription Factor-1; D050813:Octamer Transcription Factor-2; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; C497692:HCFC1 protein, human; D051863:Host Cell Factor C1; D015848:Interleukin-5; D008222:Lymphokines; D050811:Octamer Transcription Factor-1; D050813:Octamer Transcription Factor-2; C494165:POU2F1 protein, human; C058435:POU2F2 protein, human; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D000001:Calcimycin; D013755:Tetradecanoylphorbol Acetate