Functional analysis of zinc finger proteins that bind to the silencer element in the glutathione transferase P gene

Glutathione transferase P (GST-P) gene expression is repressed in normal rats but markedly promoted during the early stage of chemical hepatocarcinogenesis. We have previously identified a silencer region in this gene promoter. The silencer is composed of several cis-elements to which at least three proteins (Silencer factor-A, -B, and -C: SF-A, SF-B, and SF-C) are known to bind. We cloned and characterized the nuclear factor 1 family and the CCAAT/enhancer-binding protein family as SF-A and SF-B, respectively. Recently, zinc finger proteins as candidates for SF-C. which binds to GST-P silencer 2 (GPS2), were isolated. These proteins include four Krüppel-like proteins (BTEB2, EZF, LKLF, and TIEG1) and other factors containing multiple zinc finger motifs (TFIIIA and MZFP). In the present study, we found that the zinc finger proteins showed the same DNA-binding affinities to GPS2. Moreover, transfection analyses revealed that BTEB2, EZF, and TIEG1 repressed the GST-P promoter activity. Therefore, these three factors might contribute to the repression of the GST-P gene expression in normal rat liver.