CDNA cloning and characterization of 5′ upstream promoter region of porcine mef2c gene

Abstract

The myocyte enhancer factor 2C (MEF2C) is a member of the MEF2 family of transcription factors, involved in skeletal muscle development. In this study we report the cDNA sequence and isolate the 5' upstream region of the mef2c gene from porcine genomic DNA using PCR-based GenomeWalker. The open reading frame of porcine mef2c cDNA covers 1,392 bases, encoding 464 amino acids, which show 94% identity with human MEF2C at the level of the primary protein structure. Annear the C terminus of mef2c, a 96-nt sequence appear to represent alternatively spliced transcripts was present in some cDNAs and absent in the other. No typical TATA, GC box or CAAT box binding site was found in porcine mef2c 5' upstream region, whereas some potential binding sites for MyoD (E-box), MEF2 and MBF1 were present in the proximal upstream region. Transfection of the mef2c 5' upstream region with EGFP into cos7 cells demonstrated that the region from -162 to +115 bp immediately 5' of the exon 1 was sufficient to direct strong EGFP protein expression. Co-transfection assays demonstrated that MBF1 bound the mef2c promoter and inhibited mef2c expression. These results may be useful for elucidating the regulation mechanisms of mef2c, which interacts with other factors to regulate target genes.