Characterisation of inducible DNase I hypersensitive sites flanking the human interleukin-5 gene.
Interleukin-5 (IL-5) production is necessary for eosinophilia associated with allergic conditions and parasitic infection. IL-5 mRNA is transiently expressed by activated T-lymphocytes. In this report, we have analysed DNA regulatory regions associated with inducible IL-5 expression in the human HSB-2 T-cell line. Only low levels of transcriptional activity were induced in cells transfected with up to 1.2 kb of DNA upstream of the IL-5 gene. DNase I hypersensitivity analysis was employed to identify additional regulatory sequences located outside this region. Two hypersensitivity sites (HS) were identified, one 2.5 kb 5' and the other 1.6 kb 3' from the gene, that were induced on activation of HSB-2 cells by stimuli that induced IL-5 expression. The 5' site, but not the 3' site, was found in primary human T-cells. The presence of the 5' HS did not always coincide with IL-5 expression. Inclusion of the region encompassing the 5' HS in promoter studies mediated a moderate increase in transcriptional activity, suggesting that enhancer elements essential for induction of maximal IL-5 transcription reside at a greater distance from the IL-5 gene.
Valentine
J E
JE
Centre for Immunology, University of New South Wales and St. Vincent's Hospital, Sydney, NSW 2010, Australia.
Sewell
W A
WA
D002460:Cell Line; D003850:Deoxyribonuclease I; D017930:Genes, Reporter; D006801:Humans; D015848:Interleukin-5 / Q000096:biosynthesis / Q000235:genetics*; D019169:Jurkat Cells; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D015848:Interleukin-5; D011993:Recombinant Fusion Proteins; D008156:Luciferases; D003850:Deoxyribonuclease I