Hepatocyte growth factor (HGF) is a multifunctional growth factor implicated in a variety of tissue restructuring processes. Since HGF acts as a highly potent mitogen, HGF expression is suggested to be under a well-defined transcriptional control. The 5' sequence of the HGF gene clusters a set of several binding sites for transcription factors in a so-called multiconsensus region (MCR) located between -230 and 260. Our studies demonstrate that a NF1-like element and the E-1-box of the MCR form the main complexes with nuclear proteins and that both are involved in transcriptional silencing of the HGF gene in non-HGF expressing cell types. The E-1-box of two tandemly arranged E-boxes was shown to be a binding site of high affinity interacting with the upstream stimulatory factor (USF). While recombinant expression of a wild-type USF did not affect gene expression, a USF variant lacking the DNA binding domain restored the MCR mediated transcriptional repression, In conclusion. our data provide evidence that USF is a central factor of cell-type specific HGF regulation. acting in cooperation with additional regulatory proteins as a bivalent mediator of transcriptional activation or repression. (C) 2002 Elsevier Science (USA). All rights reserved.