Human 15-lipoxygenase gene promoter: analysis and identification of DNA binding sites for IL-13-induced regulatory factors in monocytes.
In order to study the transcriptional control of 15-LO expression, we have cloned and sequenced the human 15-LO promoter region. The 15-LO promoter is associated with a CpG island at the 5'-end of the gene, and sequence analysis reveals putative Sp1 and Ap2 binding site/s and absence of TATA or CAAT motifs. Transcription is initiated at one major site. Using deletion constructs, we have defined an active promoter region of 1056 bp. Gel-shift assays revealed that transcriptional factor(s) induced only in response to IL-13 treatment of human peripheral blood monocytes bind to the 15-LO promoter DNA. Two regions, DP1 (-140 to -92 bp) and DP2 (-353 to -304 bp) of the promoter were essential for transcription in HeLa cells and human peripheral monocytes. Hela nuclear extracts contained a specific nuclear factor(s) binding to 15-LO promoter DNA which are distinct from those derived from IL-13-treated human peripheral monocyte nuclear extracts. In addition, fluorescent in situ hybridization (FISH) results refined the previous localization of 15-LO to human chromosome 17p13.3.
Center for Glomerulonephritis, Renal Division Emory University, and Veterans Affairs Medical Center, Atlanta, GA 30033, USA.
U
U
Kelavkar
S
S
Wang
A
A
Montero
J
J
Murtagh
K
K
Shah
K
K
Badr
D000818:Animals; D001093:Arachidonate 15-Lipoxygenase / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D002886:Chromosomes, Human, Pair 17; D004247:DNA / Q000378:metabolism; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D018793:Interleukin-13 / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011817:Rabbits; D016415:Sequence Alignment; D014157:Transcription Factors / Q000378:metabolism; D015533:Transcriptional Activation
D004268:DNA-Binding Proteins; D018793:Interleukin-13; D014157:Transcription Factors; D004247:DNA; D001093:Arachidonate 15-Lipoxygenase