The bifactorial endosperm box of γ‐zein gene: characterisation and function of the Pb3 and GZMcis‐acting elements

Open Access
Publisher Website
Google Scholar
Abstract
The proximal region of the γ‐zein promoter (γ Z ) has a functional bifactorial prolamin box element containing two cis ‐acting elements, a prolamin‐box motif (Pb3) and a GCN4‐like motif (GZM). By particle bombardment of maize endosperms with 5′ deletions and internal deletions of γ Z fused to the GUS gene, we have shown that a 135 bp region containing the bifactorial element is involved in the transcriptional activation of the γ Z promoter. However, the 135 bp region was unable to activate the γ Z promoter in the absence of a 84 bp downstream sequence. Using in vivo footprinting and gel mobility shift assays with 15 DAP endosperm nuclear extracts, we have demonstrated the presence of trans ‐acting factors that interact with Pb3 and GZM target sites. Base‐substitution mutations within Pb3 and GZM decreased transcription activity of the γ Z promoter suggesting a co‐ordinated function between the two cis ‐acting elements. Two additional cis ‐motifs upstream of the bifactorial prolamin element have been identified: a motif with high homology to the AACA elements of rice glutelin genes and an AZM motif containing an ACGT core which binds nuclear proteins other than the Opaque 2 (O2).