Progression of Coronary Atherosclerosis Is Associated with a Common Genetic Variant of the Human Stromelysin-1 Promoter Which Results in Reduced Gene Expression

doi:10.1074/jbc.271.22.13055

Progression of Coronary Atherosclerosis Is Associated with a Common Genetic Variant of the Human Stromelysin-1 Promoter Which Results in Reduced Gene Expression*

From the ^‡ Division of Cardiovascular Genetics, Department of Medicine, University College London Medical School, London WC1E 6JJ, United Kingdom, the
^§ Atherosclerosis Research Unit, King Gustaf V Research Institute, Karolinska Hospital, Stockholm S-171 76, Sweden, and the
^¶ Center for Molecular Medicine and Genetics and Department of Pathology, Wayne State University School of Medicine, Detroit, Michigan 48202

^∥ To whom correspondence should be addressed. Present address:
The Wellcome Trust Centre for Human Genetics, University of Oxford, Windmill Road, Oxford OX3 7BN, United Kingdom.
Tel.: 44-01865-742441; Fax: 44-01865-742196; E-mail: adriano.henney{at}well.ox.ac.uk.

Abstract

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.

Footnotes

↵* This work was supported by Grants PG/92021, FS/95011, and RG16 from the British Heart Foundation, Grant 8691 from the Swedish Medical Research Council, and by the Swedish Heart-Lung Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

MMP

matrix metalloproteinase

CAT

chloramphenicol acetyltransferase

EMSA

electrophoretic mobility shift assay

PAI-1

plasminogen activator inhibitor-1

VSMC

vascular smooth muscle cell

TIMP

tissue inhibitor of metalloproteinases.
- Received November 7, 1995.
- Revision received January 25, 1996.