The transcription of the chicken cardiac myosin light chain-2 (MLC-2) promoter containing a 1.3 Kb 5'-flanking DNA segment is repressed upon co-transfection with an expression vector (pMMV) containing the proto-oncogene fos in embryonic chicken cardiac muscle cells in culture. Similar concentrations of co-transfectants containing other genes e.g. luciferase were ineffective. To identify the DNA element(s) in MLC-2 gene that responds to fos-mediated inhibition, 5'-sequential deletion mutants of MLC-2 promoter were tested in a transient transfection assay. A mutant, in which the 5' distal sequence was deleted upto -1200 bp upstream of the mRNA start site was sensitive to fos inhibition, but the mutant containing -1130 bp was not, suggesting that a fos responsive element (FRE) is located between -1130 to -1200 bp upstream of the transcription initiation site. The same FRE sequence was also responsive to fos-inhibition in chicken skeletal muscle cells as well. Since over-expression of fos is implicated in repression of myogenic process, the selective inhibition of MLC-2 promoter activity by fos and identification of FRE sequence potentially important in understanding the relationship between myogenesis and the oncoprotein-mediated signal pathway(s).
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