Expression of the rat S100A1 gene in neurons, glia, and skeletal muscle

Abstract

Using a rat S100A1 cDNA probe, S100A1 expression has been documented in rat C6 glioma cells, a cell line previously thought to express only the S 100B protein. To identify the molecular mechanisms which target S100A1 gene expression to specific cell types, the rat S100A1 gene was cloned, and functional analysis of the 5′ flanking region of the gene was performed. The rat S100A1 gene was located in an 8.5 kb BamHl genomic fragment which contained 3 exons plus 1.6 kb of 5′-upstream and 0.37 kb of 3′-downstream flanking sequence. A single transcription initiation start site and a single polyadenylation signal were identified in this gene. A number of potential regulatory consensus sequences were identified in the rat S100A1 gene including general transcription factor binding sequences (TATA box, GC box and CCAAT box), cAMP regulated sequences (CRE), skeletal muscle specific sequences (E-box and M-CAT), an S100 protein element, and a (GCT) trinucleotide repeat. Analysis of an S100A1 promoter-CAT construct by ribonuclease protection assay demonstrated that this gene is functional in three S1OOA1 expressing cell lines, C6 cells, PC12 cells and L6 cells. CAT constructs containing progressive deletions of the S100A1 promoter region revealed a positive regulatory element in skeletal muscle (L6) cells between −1600/−1081. The fact that these same sequences were negative in glial (C6) cells and neutral in neuronal (PC 12) cells suggests that this region plays a major role in targeting S 100A1 expression to specific cell types. The −/1081+ 10 region contained both positive and negative elements, some of which were cell-type specific. Thus, S100A1 expression is under complex transcriptional control which involves positive and negative elements as well as cell type specific elements.