Site controlled transgenic mice validating increased expression from human matrix metalloproteinase (MMP-1) promoter due to a naturally occurring SNP

Abstract

Matrix Metalloproteinases (MMPs) comprise a family of more than 20 members, each with the ability to degrade components of the extracellular matrix. The interstitial collagenases have the unique capacity to degrade the stromal collagens, types I, II and III, the body's most abundant proteins. These collagenases include MMP-1, MMP-8, MMP-13 and MMP-14. MMP-1, with a very broad expression pattern, has major roles in mediating matrix destruction in many diseases. We have described a single nucleotide polymorphism (SNP) in the MMP-1 promoter that augments transcription. This SNP is the presence or absence of an extra guanine (G) at -1607 bp, which creates the sequence 5'-GGAA-3'(2G allele), and which is an ETS binding site. Compared to the 1G allele (5'-GAA-3'), the 2G SNP is associated with enhanced transcription of MMP-1 and increased enzymatic activity.Although murine systems are often used to model human diseases, mice have only distant homologues of human MMP-1. Therefore, we used a technique for the targeted insertion of a single copy of a gene at the HPRT locus to compare expression of the 1G and 2G alleles. We generated transgenic mice with -4372 bp of the human MMP-1 promoter containing either the 1G or 2G SNP in front of the Lac Z (E.coli ß-galactosidase) gene. We measured relative expression of the transgenes in vitro in embryonic stem (ES) cells and in fibroblasts derived from embryonic mice. Our data show modest constitutive expression of ß-galactosidase mRNA and protein from these alleles, with the 2G allele more transcriptionally active than the 1G allele. We conclude that these mice represent a model for integration of a single copy of the human MMP-1 promoter into the murine genome, and could be used to study MMP-1 gene expression in a murine system.

Association of matrix metalloproteinase gene polymorphism with temporomandibular joint degeneration

“…This binding site is close to an AP‐1 site, promoting a 37‐fold increase in transcription activity, in vitro (12). A recent study of the same group, using transgenic mice, validated the increased expression of the MMP1 2G allele in vivo (27).…”

Matrix metalloproteinase genes on chromosome 11q22 and the risk of anterior cruciate ligament (ACL) rupture

“…The MMP1 rs1799750 and MMP12 rs2276109 variants investigated in this study are promoter polymorphisms and have been shown to have functional effects (Rutter et al, 1998; Jormsjo et al, 2000; Coon et al, 2009). The 2G allele of the MMP1 rs1799750 variant creates an Ets binding site (5′‐GGA‐3′) and it has been shown to have significantly higher transcription of the MMP1 gene, when compared with the 1G allele (Rutter et al, 1998).…”

Association of a matrix metallopeptidase 1 gene polymorphism with long-term outcome of thoracic aortic aneurysm

“…The level of MMP1 expression is influenced by polymorphism in the promoter region of MMP1 (22). Such polymorphism affects both transcription of MMP1 and enzyme activity (23). Indeed, the -340A→G polymorphism (rs514921) of MMP1 affects the binding of transcription factors and influences the level of MMP1 expression (22,24).…”