Progression of coronary atherosclerosis is associated with a common genetic variant of the human stromelysin-1 promoter which results in reduced gene expression.
There is a common polymorphism in the promoter sequence of the human stromelysin-1 gene, with one allele having a run of six adenosines (6A) and the other five adenosines (5A). We have previously reported, in a 3-year follow-up study of patients with coronary atherosclerosis, that those patients who are homozygous for the 6A allele show a more rapid progression of the disease. In this study, we have investigated whether the 5A/6A promoter polymorphism plays a role in the regulation of stromelysin-1 gene expression. In transient transfection experiments, a stromelysin-1 promoter construct with 6A at the polymorphic site was found to express less of the chloramphenicol acetyltransferase reporter gene than a construct containing 5A. Electrophoretic mobility shift assay and DNase I footprinting revealed the interaction of one or more nuclear protein(s) with the DNA sequence at the 5A/6A polymorphic site. The binding of one of the nucleoprotein factors was more readily detectable with an oligonucleotide probe corresponding to the 6A allele as compared with a probe corresponding to the 5A allele. Replacing the core binding sequence with a random DNA sequence abolished the interaction between the nuclear protein(s) and the probe and also increased reporter gene expression in transiently transfected cells. Thus, the common 5A/6A polymorphism of the human stromelysin-1 promoter appears to play an important role in regulating stromelysin-1 gene expression and may be involved in the progression of coronary heart disease.
Ye
S
S
Division of Cardiovascular Genetics, Department of Medicine, University College London Medical School, London WC1E 6JJ, United Kingdom.
Eriksson
P
P
Hamsten
A
A
Kurkinen
M
M
Humphries
S E
SE
Henney
A M
AM
D000483:Alleles; D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003324:Coronary Artery Disease / Q000235:genetics* / Q000503:physiopathology*; D004247:DNA; D018983:DNA Footprinting; D005786:Gene Expression Regulation; D006801:Humans; D015810:Linkage Disequilibrium; D019278:Matrix Metalloproteinase 3; D008666:Metalloendopeptidases / Q000235:genetics*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D018807:Polymorphism, Single-Stranded Conformational; D011401:Promoter Regions, Genetic; D011485:Protein Binding
D009687:Nuclear Proteins; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3