Functional analysis of the -300 region of maize zein genes.

A 43 bp fragment containing the -300 region upstream control element common to the endosperm expressed zein genes of Zea mays L. has been analyzed by in vivo and in vitro techniques. Transient transformation studies with protoplasts from a maize endosperm culture indicate that the element positively affects CaMV 35S promoter-driven gene expression, and that this effect is both orientation- and position-dependent. Band-shift and Southwestern blotting experiments demonstrate that the element is specifically bound by different sets of DNA-binding proteins from seedling and endosperm nuclei. A 2 bp substitution within the most conserved region of the element both reduces the stimulatory effect on transcription and alters the binding of nuclear proteins from both tissues.