Identification of Novel, Functional Genetic Variants in the Human Matrix Metalloproteinase-2 Gene

Abstract

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 human MMP-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 3 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 influences MMP-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.

Clinical impact of MMP and TIMP gene polymorphisms in gastric cancer

“…The (À1306C4T) SNP in the promoter of the MMP-2 gene has also been found to diminish promoter activity by abolishing the Sp1-binding site (Price et al, 2001). Consequently, the variant genotypes (CT/TT) are expected to produce less MMP-2 antigen, which consequently might be associated with decreased cancer risk or better survival of the patients (Sier et al, 1996).…”

Association between the -1306C/T polymorphism of matrix metalloproteinase-2 gene and lumbar disc disease in Chinese young adults

“…Recently, Price et al reported a functional SNP of the MMP-2 gene [14]. The -1306C?T transition in the promoter region of MMP-2 disrupts an Sp1-binding site (CCACC box), leading to a strikingly lower promoter activity with the T allele.…”

“…MMP-2 genotypes were determined by PCR and direct DNA sequencing. Oligonucleotide primers (forward, 5 0 -CTG ACC CCC AGT CCT ATC TGC C-3 0 ; reverse, 5 0 -TGT TGG GAA CGC CTG ACT TCA G-3 0 ) were used to amplify genomic DNA fragments [14]. PCR was accomplished in a total volume of 25 ll containing 100 ng DNA, 1.0 lM each primer, 0.2 mM dNTP, 2.0 mM MgCl 2 , 1.0 U Taq DNA polymerase with 1· reaction buffer and 2% dimethylsulfoxide.…”

“…The C?T transition at -1306 disrupts Sp1-binding site and results in decreased transcriptional activity, whereas the presence of the Sp1 promoter site in the -1306C allele may enhance transcription [14]. Therefore, MMP-2 protein expression would be higher in individuals who carry the CC genotype than those who carry the TT or CT genotype.…”

Association between matrix metalloproteinase (MMP)-2 polymorphisms and MMP-2 levels in hypertensive disorders of pregnancy

“…Experimental and Molecular Pathology 92 (2012) 217-221 MMP-2 activity is regulated at different levels including its transcription, activation of its latent form, and endogenous inhibition mainly by the tissue inhibitor of metalloproteinase (TIMP)-2 (Lambert et al, 2004). While two functional genetic polymorphisms in the promoter region of MMP-2 gene (g.-1306 C > T and g.-735 C > T) may affect MMP-2 expression and activity (Price et al, 2001;Yu et al, 2004), and therefore promote cardiovascular diseases (Lacchini et al, in press;Mizon-Gerard et al, 2004), no previous study has studied MMP-2 gene polymorphisms in hypertensive disorders of pregnancy.…”