Transcription factor Ets-1 regulates gelatinase a gene expression in mesangial cells

Ets transcription factors are involved in cell growth and angiogenesis. Ets-1 targets include members of the matrix metalloproteinase superfamily. In inflammatory glomerular diseases, the patterns and regulation of Ets expression have not been fully characterized. In the present study, nuclear binding activities to the consensus Ets-1/PEA3 motif were detected in mesangial cells (MC), and the Ets-1 protein was positively identified by Western blotting, reverse transcription PCR (RT-PCR), and DNA-binding studies. The 5' flanking regions of the human and rat gelatinase A genes contain clusters of potential Ets-1 binding motifs, one of which is evolutionarily conserved. Using a series of 5' deletion reporter constructs of the rat gelatinase A gene and an Ets-1 expression plasmid, a concentration-dependent threefold trans-activation of gene expression mapped to the conserved Ets-1 binding motif at -1004/-1053 bps, designated responsive element-2 (RE-2). The RE-2 was operative within the context of the homologous gelatinase A promoter but not with a heterologous simian virus 40 promoter. Specific Ets-1 binding to this sequence was demonstrated by DNA-binding studies. Transient expression of an Ets-1 expression plasmid increased gelatinase A protein expression. Our findings identify an additional matrix metalloproteinase family member, gelatinase A, as an Ets-1 responsive gene in MC that may play a role in the high level expression of this enzyme in inflammatory glomerular diseases.