GENES: STRUCTURE AND REGULATION
Identification of Novel, Functional Genetic Variants in the Human Matrix Metalloproteinase-2 Gene: ROLE OF Sp1 IN ALLELE-SPECIFIC TRANSCRIPTIONAL REGULATION*

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Matrix metalloproteinase-2 (MMP-2) is an enzyme with proteolytic activity against matrix and nonmatrix proteins, particularly basement membrane constituents. Thus, any naturally occurring genetic variants that directly affect gene expression and/or protein function would be expected to impact on progression of pathological processes involving tissue remodeling. We scanned a 2-kilobase pair promoter region and all 13 exons of the humanMMP-2 gene, from a panel of 32 individuals, and we identified the position, nature, and relative allele frequencies of 15 variant loci as follows: 6 in the promoter, 1 in the 5′-untranslated region, 6 in the coding region, 1 in intronic sequence, and 1 in the 3′-untranslated region. The majority of coding region polymorphisms resulted in synonymous substitutions, whereas three promoter variants (at −1306, −790, and +220) mapped onto cis-acting elements. We functionally characterized all promoter variants by transient transfection experiments with 293, RAW264.7, and A10 cells. The common C → T transition at −1306 (allele frequency 0.26), which disrupts an Sp1-type promoter site (CCACC box), displayed a strikingly lower promoter activity with the T allele. Electrophoretic mobility shift assays confirmed that these differences in allelic expression were attributable to abolition of Sp1 binding. These data suggest that this common functional genetic variant influencesMMP-2 gene transcription in an allele-specific manner and is therefore an important candidate to test for association in a wide spectrum of pathologies for which a role for MMP-2 is implicated, including atherogenesis and tumor invasion and metastasis.AJ298926

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Published, JBC Papers in Press, December 12, 2000 DOI 10.1074/jbc.M010242200

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This work was supported in part by British Heart Foundation Grant RG/1995008.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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) .

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Recipient of a British Heart Foundation Studentship Grant FS/97051.

British Heart Foundation Basic Science Lecturer and supported by Grant BS/99003.