Functional identification of the transcriptional regulatory elements within the promoter region of the human ventricular myosin alkali light chain gene.

摘要: Abstract We have identified and functionally characterized DNA sequences that regulate the expression of human ventricular/slow twitch isoform myosin alkali light chain (VLC1) gene. By using primer extension S1 nuclease mapping techniques, we shown VLC1 gene is transcribed from identical site in ventricular slow skeletal muscles. Comparison +1 to -1296 genes for mouse showed 5'-proximal flanking region, up about 220 nucleotides, was highly conserved (83% homology). To determine location sites may be important function promoter, a series transient vectors containing progressive deletions 5'-flanking sequence fused bacterial chloramphenicol acetyltransferase (CAT) introduced into myogenic nonmyogenic cells. Deletion mutagenesis between -357 +40 revealed presence positive negative activity all cells tested. demonstrated minimal promoter required generate muscle cell-specific region -94 -64 upstream cap element located -107 found effect both These two proximal regions appear act together cell type-specific high level Competition gel retardation assays CArG -96 -87 interacts specifically with nuclear extracts compete binding present cardiac alpha-actin serum response c-fos results strongly suggested similar, if not identical, box proteins interact different VLC1, alpha-actin, genes.