Isolation of novel, transcriptionally active AP-1 binding sites: implications for cellular transformation.

Increased AP-1 DNA-binding activity, in the context of TRE-binding, is not a consequence of Fos transformation. In this report we investigate the possibility of a change in binding site preference by vFosAP-1 compared with AP-1 from an untransformed cell. Fos binding sites were immunoselected from random sequence oligonucleotides using a pan Fos anti-serum with nuclear protein from quiescent FBRp75v-fos-transformed (FBR) and normal (208F) rat fibroblasts. The selected oligonucleotides were aligned by computer and a consensus described for the sequences bound by AP-1 from the two cell lines. The vFos binding site is shown to be a consensus TRE, whereas the sequence ACCACATC is described as the cellular Fos protein family consensus. We demonstrate that sequences differing from the TRE consensus can bind AP-1 and direct transcription. AP-1 DNA-binding activity differs between normal and transformed cells with several of the selected oligonucleotides. These sequences also demonstrate differential transcriptional activation between normal and transformed cells. In particular, the 208F consensus has no transcriptional activity in FBR cells. Further, EGF differentially influences the transcriptional activity of the oligonucleotides in 208F and FBR cells. Our results suggest that AP-1 may change its preferred binding site depending on the proteins available at any given time, the sequences flanking a non-consensus TRE or even the environment in which the cell exists. These differences in binding site preference and transcriptional activation may result in the increased transforming ability of the v-fos oncogene compared with the c-fos proto-oncogene and may extend the potential target genes beyond those with an AP-1 consensus binding site.