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J. Biol. Chem., Vol. 266, Issue 13, 8511-8516, May, 1991
BP Kennedy, RE Diehl, Y Boie, M Adam and RA Dixon
The human gene for the recently identified 5-lipoxy-genase-activating
protein (FLAP) has been cloned. The gene was isolated from two different
genomic libraries and is contained within four overlapping bacteriophage
clones. The gene spans greater than 31 kilobases and consists of five small
exons and four large introns. Southern blot analysis of human genomic DNA
suggests the presence of a single FLAP gene per haploid genome. A
restriction site polymorphism was identified in intron II of the gene. This
restriction fragment length polymorphism appears to be present in the
normal population at a fairly high frequency. The transcription initiation
site was located, at an adenine residue, 74 base pairs upstream of the ATG
initiation codon. Examination of the sequence of the gene 5' to the mRNA
start site revealed the presence of a possible TATA box (TGTAAT) 22 base
pairs upstream and potential AP-2 and glucocorticoid receptor binding
sites. Functional analysis of the FLAP gene promoter was assayed by
transient transfection of mouse P388D1 cells (macrophage) and human HepG2
cells (hepatoma) with 5'-flanking sequences of the FLAP gene fused upstream
of the chloramphenicol acetyltransferase reporter gene. Expression in the
mouse macrophage cell line of the various FLAP gene promoter constructs
revealed both tissue specificity and enhancer-like activities whereas in
the hepatoma cell line only a minimum level of activity was obtained.
Gene characterization and promoter analysis of the human 5-lipoxygenase- activating protein (FLAP)
Department of Molecular Biology, Merck Frosst Center for Therapeutic Research, Pointe Claire-Dorval, Quebec, Canada.
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