The two allele sequences of a common polymorphism in the promoter of the plasminogen activator inhibitor-1 (PAI-1) gene respond differently to interleukin-1 in HepG2 cells.
We have detected a common (allele frequency 0.53/0.47) single base pair insertion/deletion polymorphism 675 base pairs upstream from the start of transcription of the plasminogen activator inhibitor-1 (PAI-1) gene, using the chemical cleavage mismatch analysis. "Band shift assays" suggest that the allele with the single base pair insertion contains an additional protein binding site which is not present in the del allele. Competition experiments confirmed that the binding was specific to the sequence of the ins allele and suggest that proteins bound to this site may be NF-kB-like proteins. Analysis of chloramphenicol acetyltransferase (CAT) mRNA produced by constructs containing the PAI-1 promoter (-805 to +83) showed that the deletion allele produced six times more mRNA than the insertion allele in response to interleukin-1 (p < 0.001). In a sample of 107 young patients with previous myocardial infarction and 95 healthy population-based subjects, the del allele was associated with increased PAI-1 levels, 21% higher than the sample mean in the del homozygotes (p < 0.05). This study also suggested that individuals homozygous for the del allele may have an altered response to the acute phase stimulus. Taken together these results suggest that the insertion/deletion polymorphism in the PAI-1 promoter is of functional importance in regulating the expression of the PAI-1 gene.
Charing Cross Sunley Research Center, Hammersmith, London.
S J
SJ
Dawson
B
B
Wiman
A
A
Hamsten
F
F
Green
S
S
Humphries
A M
AM
Henney
D000328:Adult; D000483:Alleles; D001483:Base Sequence; D001667:Binding, Competitive; D006528:Carcinoma, Hepatocellular; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D004251:DNA Transposable Elements; D005091:Exons; D005838:Genotype; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D015850:Interleukin-6 / Q000494:pharmacology; D008113:Liver Neoplasms; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009203:Myocardial Infarction / Q000235:genetics; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D009838:Oligodeoxyribonucleotides; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012016:Reference Values; D017384:Sequence Deletion; D014162:Transfection; D014407:Tumor Cells, Cultured
D004251:DNA Transposable Elements; D007375:Interleukin-1; D015850:Interleukin-6; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D015500:Chloramphenicol O-Acetyltransferase