Molecular dissection of the interaction between the transcriptional activator ZmMRP-1 and the promoter of BETL-1

The interaction between the transfer cell specific transcriptional activator ZmMRP-1 and the promoter of the transfer cell specific gene BETL-1 constitutes an exceptionally robust system. Reporter constructs containing the BETL-1 promoter are virtually silent in a variety of cell types, from maize leaves to yeast. The introduction of ZmMRP-1 in co-transformation assays leads to the transactivation of the reporter construct by up to two orders of magnitude. In this work we have investigated the molecular basis of this interaction. We found that the BETL-1 promoter includes four potential targets for ZmMRP-1 binding, consisting of a 12 bp motif containing two repeats. Co-transformation assays and electrophoretic mobility shift experiments identified the sequence TATCTCTATCTC as the preferred one for the interaction with the transcription factor. Identification of similar sequences in other transfer cell specific promoters lead us to propose as a transfer cell box a sequence related to those identified in the BETL-1 promoter, positioned 50-100 bp upstream the TATA box.