Display Settings:


Send to:

Choose Destination
J Biol Chem. 1997 Feb 21;272(8):5219-28.


Author information

  • 1Department of Pathology, Anatomy, and Cell Biology,Jefferson Medical C ollege, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA. Iozzo@lac.jci.tju.edu


Perlecan, a modular heparan sulfate proteoglycan of basement membranes and cell surfaces, plays a crucial role in regulating the assembly of extracellular matrices and the binding of nutrients and growth factors to target cells. To achieve a molecular understanding of perlecan gene regulation, we isolated the 5'-flanking region and investigated its functional promoter activity and its response to cytokines. Transient cell transfection assays, using plasmid constructs harboring the perlecan promoter linked to the chloramphenicol acetyltransferase reporter gene, demonstrated that the largest approximately 2.5-kilobase construct contained maximal promoter activity. This promoter region was functionally active in a variety of cells of diverse histogenetic origin, thus corroborating the widespread expression of this gene product. Stepwise 5' deletion analyses demonstrated that the -461-base pair (bp) proximal promoter retained approximately 90% of the total activity, and internal deletions confirmed that the most proximal sequence was essential for proper promoter activity. Nanomolar amounts of transforming growth factor-beta induced 2-3-fold perlecan mRNA and protein core levels in normal human skin fibroblasts, and this induction was transcriptionally regulated; in contrast, tumor necrosis factor-alpha had no effect and was incapable of counteracting the effects of TGF-beta. Using additional 5' deletions and DNase footprinting analyses, we mapped the TGF-beta responsive region to a sequence of 177 bp contained between -461 and -285. This region harbored a 14-bp element similar to a TGF-beta-responsive element present in the promoters of collagen alpha1(I), alpha2(I), elastin, and growth hormone. Electrophoretic mobility shift assays and mutational analyses demonstrated that the perlecan TGF-beta-responsive element bound specifically to TGF-beta-inducible nuclear proteins with high affinity for NF-1 member(s) of transcription factors.

[PubMed - indexed for MEDLINE]
Free full text
PubMed Commons home

PubMed Commons

How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for HighWire
    Loading ...
    Write to the Help Desk