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M001322200v1
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Papers In Press, published online ahead of print April 12, 2000
J. Biol. Chem, 10.1074/jbc.M001322200
Submitted on February 16, 2000
Accepted on April 11, 2000

The hematopoietic transcription factor PU.1 represses gelatinase A transcription in glomerular mesangial cells

Sigrid Harendza, David H. Lovett, and Rolf A.K. Stahl

Medizinische Klinik, Universitats-Krankenhaus Eppendorf, D-20246, Hamburg

Corresponding Author: harendza{at}uke.uni-hamburg.de

The matrix metalloproteinase gelatinase A plays a key role in the evolution of glomerular injury and is a major contributing factor to the development of glomerulosclerosis. Prior studies have focused on a potent cis-acting enhancer element located in the near 5? flanking region of the rat and human gelatinase A genes (1-3). Given the combinatorial nature of transcriptional regulation we examined additional regions of the 5? flanking region of the rat gelatinase A gene to identify further regulatory elements. In this study the identification of a silencing element located between ?1903 and ?1847 bp of the 5? flanking region of the rat gelatinase A gene is reported. Sequence analysis, electrophoretic mobility studies and transfection experiments demonstrate that a specific binding sequence for the hematopoietic transcription factor PU.1 is present within the silencing sequence. PU.1 activity is absolutely required for the expression of silencing activity within the context of transfected glomerular mesangial cells. Western blots identify the PU.1 protein within nuclear extracts of mesangial cells, and co-transfection with a PU.1 expression vector directly augments silencing activity. These studies underscore the complex patterns of gelatinase A transcriptional regulation and also strongly suggest that glomerular mesangial cells are ultimately derived from bone marrow cells.


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