Stromelysin-1 expression is activated in vivo by Ets-1 through palindromic head-to-head Ets binding sites present in the promoter

Abstract

Regulation of the gene expression of Stromelysin-1 (matrix metalloproteinase-3), a member of the matrix metalloproteinase family, is critical for tissue homeostasis. The Stromelysin-1 promoter is known to be transactivated by Ets proteins through palindromic headto-head Ets binding sites (EBS), an unusual configuration among metalloproteinase promoters. Patterns of increased co-expression of Stromelysin-1 and Ets-1 genes have been observed in pathological processes such as rheumatoid arthritis, glomerulonephritis and tumor invasion. In this context, we show in a synovial fibroblastic model cell line (HIG-82), which is able to co-express Stromelysin-1 and Ets-1, that the EBS palindrome is essential for the expression of Stromelysin-1. More precisely, using electrophoretic mobility shift assays, DNA affinity purification and chromatin immunoprecipitation, we demonstrate that endogenous Ets-1, but not Ets-2, is present on this palindrome. The use of a dominant-negative form of Ets-1 and the decrease of Ets-1 amount either by fumagillin, an antiangiogenic compound, or by short interfering RNA show that the activation rate of the promoter and the expression of Stromelysin-1 correlate with the level of endogenous Ets-1. Thus, it is the first demonstration, using this cellular model, that endogenously expressed Ets-1 is actually a main activator of the Stromelysin-1 promoter through its effective binding to the EBS palindrome.

Sub-circuits of a gene regulatory network control a developmental epithelial-mesenchymal transition

“…However, several TF knockdowns that successfully degraded laminin in our model system have been shown to promote basement membrane remodeling in other systems. Previous studies have demonstrated that ets1 directly upregulates mmp1 (collagenase 1), mmp3 (stromelysin 1), mmp9 (gelatinase B) and mmp13 (collagenase 3) (Westermarck et al, 1997;Baillat et al, 2006;Ghosh et al, 2012). Snail and twist are also associated with upregulated MMPs (Sánchez-Tilló et al, 2012), although this evidence is indirect.…”

Interferon‐γ regulation of TNFα‐induced matrix metalloproteinase 3 expression and migration of human glioma T98G cells

“…Previous studies on TNFa-dependent MMP-3 activation have identified Ets-1 and NFjB as the main effectors of gene expression in synovial fibroblastic cells 16 and monocyte-derived macrophages. 17 In this report, we have made similar observations on MMP-3 induction in the T98G cells upon TNFa treatment.…”

“…[15][16][17] It has been shown that Ets-1 and NFjB are the main transcription factors in controlling MMP-3 gene expression inducible by TNFa in fibroblasts and macrophages. 16,17 To elucidate the underlying mechanisms by which IFNg suppresses TNFa-induced MMP-3 expression, we investigated the effects of IFNg on the DNA-binding activity of Ets-1, NFjB and AP-1 to their respective enhancer elements. Results of EMSA suggested that TNFa activated DNA-binding of Ets-1 and NFjB to their respective oligodeoxynucleotide enhancer sequences (Fig.…”

Ets-1 p27: a novel Ets-1 isoform with dominant-negative effects on the transcriptional properties and the subcellular localization of Ets-1 p51

“…Western blot analysis was performed as previously described (Baillat et al, 2006) using (1) 30 mg of total cell lysate proteins or (2) 25 ml of total cell lysates in the case of reporter gene assays, which were boiled in Laemmli buffer (50 mM Tris (pH 6.8), 2% sodium dodecyl sulfate, 5% b-mercaptoethanol, 10% glycerol, 0.1% bromophenol blue). Primary antibodies used (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were the C-20 anti-Ets-1, C-20 anti-Ets-2, 16 anti-PEA3, C-14 anti-Erk-2, H-79 anti-cJun and H-125 anti-cFos.…”