Interacting cis elements in the plastocyanin promoter from spinach ensure regulated high-level expression

Abstract

The spinach plastocyanin promoter contains most, if not all, cis elements crucial for its activity downstream of -259 bp relative to the transcription start site. The -259/-79 bp promoter fragment is capable of conferring glucuronidase (GUS) gene expression on the minimal -90/+3 bp 35S RNA promoter of CaMV and -51/+60 bp plastocyanin promoter, regardless of its orientation. Using 5' promoter deletion analysis and site directed mutagenesis we identified three regions, designated PC-1 (-195/-188), PC-2 (-179/-164) and PC-3 (-90/-77) for promoter function. An interaction between PC-3 and the upstream elements is required for high levels of expression. All these sequences contain binding sites for protein factors, as shown by gel shift assays. PC-3 includes a binding site with some resemblance to GT-1 box II, but additional nucleotide sequences immediately downstream of this motif, which are conserved among all published plastocyanin promoters, are required as well. The sequence interval -168/-79 bp is sufficient to confer light-responsive, organ-specific and chloroplast-dependent GUS gene expression on minimal promoters.

Coordination of plastid and nuclear gene expression

“…Deletion analysis of several of these promoters has identified shorter regions that are able to direct similar patterns of expression in norflurazon-treated seedlings (Bolle et al 1994;Lü bberstedt et al 1994;Gray et al 1995;Kusnetsov et al 1996;Hahn & Kü ck 1999). However, in none of the promoters was it possible to separate the regulatory elements responding to norflurazon treatment from light-regulatory elements.…”

Ancestral Multipartite Units in Light-Responsive Plant Promoters Have Structural Features Correlating with Specific Phototransduction Pathways

“…These are the tobacco rbcS gene 52-bp I-G region (Argiiello-Astorga and Herrera-Estrella, 1995), a 56-bp DNA segment from the maize C4ppdkZml (encoding pyruvate, Pi dikinase) gene (Sheen, 1991), a 65-bp truncated pea rbcS 3A gene promoter (Kuhlemeier et al, 1989), a 72-bp region from the spinach rbcA (encoding Rubisco activase) gene promoter (Orozco and Ogren, 1993), a 78-bp segment from the Arabidopsis cab2 gene (Anderson et al, 1994), and an 89-bp region from the spinach plastocyanin gene promoter (Lubberstedt et al, 199413; Fig. 3) (Ha and An, 1988).…”

“…Plant lineages in which identified CMAs were found are listed, and one specific CMA-containing region where an LRE has been experimentally identified is also indicated. #, Light- Lubberstedt et al, 199413. "DREP module" is a conserved sequence including the AAAAT motif, which is critical for functional interactions between AtpC promoter and a putative in-dark repressor (Bolle et al, 1996).…”

Promoters from Genes for Plastid Proteins Possess Regions with Different Sensitivities toward Red and Blue Light

“…At the other end of the pathway(s), a variety of cis-regulatory elements have been identified that specifically interact with families of related proteins (cf. Dehesh et al, 1990;Gilmartin et al, 1990;Schindler and Cashmore, 1990;Liibberstedt et al, 1994). GT-1-binding sites, GPu(T/A)AA(T/A), for example, have been found in the upstream regions of various light-regulated genes (Stockhaus et al, 1987;Manzara and Gruissem, 1988;Kay et al, 1989;Dehesh et al, 1990;Liibberstedt et al, 1994).…”

“…Dehesh et al, 1990;Gilmartin et al, 1990;Schindler and Cashmore, 1990;Liibberstedt et al, 1994). GT-1-binding sites, GPu(T/A)AA(T/A), for example, have been found in the upstream regions of various light-regulated genes (Stockhaus et al, 1987;Manzara and Gruissem, 1988;Kay et al, 1989;Dehesh et al, 1990;Liibberstedt et al, 1994). The core element of the G-box sequence (CACGTG), in tum, is not only present in promoters of lightregulated genes (Giuliano et al, 1988;Schulze-Lefert et al, 1989;Staiger et al, 1989;Donald and Cashmore, 1990;Weisshaar et al, 1991), but also in promoters that are regulated by other stimuli (cf.…”