Linked Common Polymorphisms in the Gelatinase A Promoter Are Associated with Diminished Transcriptional Response to Estrogen and Genetic Fitness

Abstract

Gelatinase A (matrix metalloproteinase-2) plays a prominent role in multiple biologic processes. Prior studies have established critical roles for gelatinase A transcriptional regulation by defined enhancer elements. To determine possible functional single nucleotide polymorphisms within these elements, we determined the single nucleotide polymorphism distribution within 1,665 bp of the gelatinase A 5 -flanking region, using a healthy homogeneous Caucasian study group of 463 individuals. Among the polymorphisms detected, a G 3 A transition at bp ؊1575 was located immediately 5 to a half-palindromic potential estrogen receptor binding site. In estrogen receptor-positive MCF-7 cells the ؊1575G allele functioned as an enhancer, whereas the ؊1575A allele reduced transcription activity significantly. Gel shift assays confirmed that the differences in allelic expression affected binding of the estrogen receptor-␣ to this region. Cotransfection experiments with an estrogen receptor-␣ expression vector in MDA-MB-231 cells, which do not constitutively express an estrogen receptor, revealed that estrogen receptor is absolutely required for enhancing activity. Allelic distribution analysis indicated that a previously reported C 3 T transition within an Sp1 binding site at ؊1306 was in linkage disequilibrium with the ؊1575G 3 A transition. Luciferase reporter studies of the linked variant ؊1575A ؊1306T allele versus the wild type ؊1575G ؊1306C allele demonstrated an additive reduction in estrogen-dependent reporter activity. The frequency of the ؊1575G 3 A transition deviated significantly from the expected Hardy-Weinberg distribution in two independently assembled study populations consisting of healthy adult blood donors and newborns of Caucasian origin, both with a calculated 21% reduction in genetic fitness. Gelatinase A is a known estrogen-responsive gene and the demonstration of a loss of function polymorphism within an operational estrogen receptor binding site associated with a decrease in genetic fitness underscores the biologic significance of promoter polymorphism analyses.The individual components of the relatively large matrix metalloproteinase gene family play multiple roles in the interaction of cells with their surrounding extracellular matrix. As such, this family of evolutionarily conserved enzymes is intimately involved in the diverse physiologic processes that make up normal growth and development, as well as neoplasia, metastasis, angiogenesis, and tissue fibrosis. One member of the matrix metalloproteinase gene family, gelatinase A or matrix metalloproteinase-2, has received considerable attention as a mediator of tumor angiogenesis and metastatic capability (1-3). In addition to the role of gelatinase A in malignancy, this enzyme has an important function in the normal growth and development of soft tissues. For example, gelatinase A-deficient mice develop normally but have significantly retarded growth rates (4) and diminished corneal and tumor neoangiogenesis (5). One form of the inherited osteolysi...

Subtype specific estrogen receptor action protects against changes in MMP-2 activation in mouse retinal pigmented epithelial cells

“…In RPE cell lines, transient transfection studies showed decreased baseline promoter activity when the deletion was present compared to the intact promoter. This has been noted previously for breast cancer cell lines (Harendza et al, 2003). Importantly, the E 2 mediated luciferase increase was abolished when the Sp1 site was deleted from the promoter construct.…”

“…Sp1 has been shown to be important for transcription of the MMP-2 and MMP-14 genes (Alfonso-Jaume et al, 2004;Harendza et al, 2003). Therefore we utilized an MMP-2 promoter construct containing a common functional polymorphism (−1306C → T) leading to a deletion of an Sp1 site (Price et al, 2001).…”

Matrix metalloproteinases and gastrointestinal cancers: Impacts of dietary antioxidants

“…The common C>T transition at -1306 disrupts a Sp1-type promoter site (CCACC box), leading to lower promoter activity with the T allele [54] . On the other hand, G to A substitution at the MMP-2 -1575 site reduces gene expression due to a reduction of estrogen receptor-α binding to A allele [55] . Fruh et al [56] found the presence of CC allele at MMP-2 -1306 position in H. pylori infected individuals which provide protection against esophagus adenocarcinoma.…”

Novel Functional Single Nucleotide Polymorphisms in the Latent Transforming Growth Factor-β Binding Protein-1L Promoter

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“…[35][36][37][38][39] Luciferase reporter assays revealed that the G-A haplotype caused transcriptional activation more efficiently than other haplotypes. The LTBP-1L promoter has a high GC content, but lacks a TATA box.…”