Identification of cis-regulatory elements in the myelin proteolipid protein (PLP) gene.
Regulatory elements of the proteolipid protein (PLP) gene were identified physically by footprinting and gel mobility shift assays and functionally by transfecting glial cell lines with PLP-chloramphenicol acetyltransferase chimeric genes. In both human and rat glial cells, only several hundred base pairs of upstream sequence were sufficient for high level activity of the human PLP promoter. This region contains five sites that contact nuclear proteins in vitro. More distal recognition sites may exist, as regions upstream of -524 displayed silencing activity indicative of a negative regulatory element. A series of site directed mutations revealed one essential positive element (ATGGA at -118) which is found in other genes encoding myelin proteins. Our combined biochemical and functional analyses indicate that the key cis sites for maximal tissue-specific expression of PLP in cultured glial cells are clustered near the promoter. Within this cluster are several conserved motifs that may coordinate the regulation of myelin-specific genes.
Berndt
J A
JA
Laboratory of Viral and Molecular Pathogenesis, National Institute of Neurologic Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892.
Kim
J G
JG
Hudson
L D
LD
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002678:Chimera; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D016172:DNA Fingerprinting; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D008745:Methylation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009185:Myelin Proteins / Q000235:genetics*; D018991:Myelin Proteolipid Protein; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D014162:Transfection
D009185:Myelin Proteins; D018991:Myelin Proteolipid Protein; D015500:Chloramphenicol O-Acetyltransferase