Identification of highly conserved regulatory domains and protein-binding sites in the promoters of the rat and human genes encoding the stress-inducible 78-kilodalton glucose-regulated protein.

Abstract

The gene encoding GRP78 has been shown to be constitutively expressed in many cell types and is inducible by the calcium ionophore A23187. To understand the regulation of GRP78 transcription, we analyzed the components that control its basal-level expression. By transfecting deletions into cells, we have identified a 54-nucleotide cis-acting regulatory element important for high basal-level expression and a contiguous 50-nucleotide element for both basal-level expression and A23187 induction. Using DNase footprinting assays with both rat and human GRP78 promoters, we demonstrated that the protein factors present in the HeLa cell nuclear extracts bind to the regulatory regions identified by the deletion studies. This domain contains a palindromic sequence and is highly conserved among GRP genes in Caenorhabditis elegans, chicks, rats, and humans.

Induction of internucleosomal DNA fragmentation by carcinogenic chromate: relationship to DNA damage, genotoxicity, and inhibition of macromolecular synthesis.

“…Potentially therefore, DNA-protein cross-links formed within the GRP78 promoter may inhibit the action of transacting factors required for gene induction. In fact, a number of transcription-factor binding sites, which are required for inducible gene expression, have been identified within the promoter region of the GRP78 gene (51)(52)(53)(54). DNAprotein cross-links occurred predominantly in the nuclear matrix fraction after treatment with 150 pM chromate (Table 1); and repair of these cross-links 24 hr after treatment correlated with recovery of GRP78 induction ( Figure 5; Table 1, respectively).…”

A stress response pathway from the endoplasmic reticulum to the nucleus requires a novel bifunctional protein kinase/endoribonuclease (Ire1p) in mammalian cells

“…A conserved promoter region, the glucose-regulated core sequence, in several mammalian genes encoding for ER proteins was identified as a potential cis-acting regulatory element equivalent to the yeast UPRE (Resendez et al 1988). Despite the sequence similarity between the mammalian glucose-regulated core sequence and the S. cerevisiae UPRE, no single element in this promoter region appears necessary and sufficient to mediate transcriptional induction as described for the UPRE in yeast cells.…”

Genomic organization and full‐length cDNA sequence of human collagen X

“…A part of this sequence (nucleotide number -83 to -72 (Fig. 2) resembles the binding motif of transcription factors of the Ets family of protooncogenes which bind to a purine-rich motif around a conserved GAG trinucleotide [38]. AT-rich motifs at a distance of about 100 nucleotides to the transcription initiation site have also been found in human [39] and rat [40] and identified as binding sites for HNF-1 respectively Pit-1.…”