title: Naturally occurring mutations in the human 5-lipoxygenase gene promoter that modify transcription factor binding and reporter gene transcription.
abstract: Five lipoxygenase (5-LO) is the first committed enzyme in the metabolic pathway leading to the synthesis of the leukotrienes. We examined genomic DNA isolated from 25 normal subjects and 31 patients with asthma (6 of whom had aspirin-sensitive asthma) for mutations in the known transcription factor binding regions and the protein encoding region of the 5-LO gene. A family of mutations in the G + C-rich transcription factor binding region was identified consisting of the deletion of one, deletion of two, or addition of one zinc finger (Sp1/Egr-1) binding sites in the region 176 to 147 bp upstream from the ATG translation start site where there are normally 5 Sp1 binding motifs in tandem. Reporter gene activity directed by any of the mutant forms of the transcription factor binding region was significantly (P < 0.05) less effective than the activity driven by the wild type transcription factor binding region. Electrophoretic mobility shift assays (EMSAs) demonstrated the capacity of wild type and mutant transcription factor binding regions to bind nuclear extracts from human umbilical vein endothelial cells (HUVECs). These data are consistent with a family of mutations in the 5-LO gene that can modify reporter gene transcription possibly through differences in Sp1 and Egr-1 transactivation.
authors.affiliation: Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.
authors.forename: K H
authors.initials: KH
authors.lastname: In
authors.forename: K
authors.initials: K
authors.lastname: Asano
authors.forename: D
authors.initials: D
authors.lastname: Beier
authors.forename: J
authors.initials: J
authors.lastname: Grobholz
authors.forename: P W
authors.initials: PW
authors.lastname: Finn
authors.forename: E K
authors.initials: EK
authors.lastname: Silverman
authors.forename: E S
authors.initials: ES
authors.lastname: Silverman
authors.forename: T
authors.initials: T
authors.lastname: Collins
authors.forename: A R
authors.initials: AR
authors.lastname: Fischer
authors.forename: T P
authors.initials: TP
authors.lastname: Keith
authors.forename: K
authors.initials: K
authors.lastname: Serino
authors.forename: S W
authors.initials: SW
authors.lastname: Kim
authors.forename: G T
authors.initials: GT
authors.lastname: De Sanctis
authors.forename: C
authors.initials: C
authors.lastname: Yandava
authors.forename: A
authors.initials: A
authors.lastname: Pillari
authors.forename: P
authors.initials: P
authors.lastname: Rubin
authors.forename: J
authors.initials: J
authors.lastname: Kemp
authors.forename: E
authors.initials: E
authors.lastname: Israel
authors.forename: W
authors.initials: W
authors.lastname: Busse
authors.forename: D
authors.initials: D
authors.lastname: Ledford
authors.forename: J J
authors.initials: JJ
authors.lastname: Murray
authors.forename: A
authors.initials: A
authors.lastname: Segal
authors.forename: D
authors.initials: D
authors.lastname: Tinkleman
authors.forename: J M
authors.initials: JM
authors.lastname: Drazen
mesh_terms: D000483:Alleles; D001249:Asthma / Q000235:genetics*; D001483:Base Sequence; D018387:Codon, Initiator; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics; D004342:Drug Hypersensitivity / Q000235:genetics; D051766:Early Growth Response Protein 1; D005091:Exons; D005260:Female; D017930:Genes, Reporter / Q000235:genetics*; D006801:Humans; D017874:Immediate-Early Proteins; D008084:Lipoxygenase / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D018807:Polymorphism, Single-Stranded Conformational; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics*
chemical_list: D018387:Codon, Initiator; D017931:DNA Primers; D004268:DNA-Binding Proteins; C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; D017874:Immediate-Early Proteins; D015682:Retroviridae Proteins, Oncogenic; D014157:Transcription Factors; D008084:Lipoxygenase