The myosin light chain enhancer and the skeletal actin promoter share a binding site for factors involved in muscle-specific gene expression.
The myosin light chain (MLC) 1/3 enhancer (MLC enhancer), identified at the 3' end of the skeletal MLC1/3 locus, contains a sequence motif that is homologous to a protein-binding site of the skeletal muscle alpha-actin promoter. Gel shift, competition, and footprint assays demonstrated that a CArG motif in the MLC enhancer binds the proteins MAPF1 and MAPF2, previously identified as factors interacting with the muscle regulatory element of the skeletal alpha-actin promoter. Transient transfection assays with constructs containing the chloramphenicol acetyltransferase reporter gene demonstrated that a 115-bp subfragment of the MLC enhancer is able to exert promoter activity when provided with a silent nonmuscle TATA box. A point mutation at the MAPF1/2-binding site interferes with factor binding and abolishes the promoter activity of the 115-bp fragment. The observation that an oligonucleotide encompassing the MAPF1/2 site of the MLC enhancer alone cannot serve as a promoter element suggests that additional factor-binding sites are necessary for this function. The finding that MAPF1 and MAPF2 recognize similar sequence motifs in two muscle genes, simultaneously activated during muscle differentiation, implies that these factors may have a role in coordinating the activation of contractile protein gene expression during myogenesis.
Ernst
H
H
Department of Medicine, Medical University of South Carolina, Charleston 29425.
Walsh
K
K
Harrison
C A
CA
Rosenthal
N
N
D000199:Actins / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D002642:Chick Embryo; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D008745:Methylation; D051379:Mice; D008969:Molecular Sequence Data; D009132:Muscles / Q000502:physiology*; D009218:Myosins / Q000235:genetics*; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism; D015345:Oligonucleotide Probes; D010750:Phosphoproteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D014162:Transfection
D000199:Actins; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D015345:Oligonucleotide Probes; D010750:Phosphoproteins; D009218:Myosins