title: Regulation of myelin basic protein-encoding gene transcription in rat oligodendrocytes.
abstract: The myelin (My) basic protein-encoding gene (MBP) is specifically expressed by oligodendrocytes (OL) in the central nervous system (CNS) and by Schwann cells in the peripheral nervous system (PNS). To define cell-type-specific regulatory elements, a series of chimaeric constructs containing varying lengths of the 5'-flanking region and exon 1 of MBP linked to the cat reporter gene was transfected into several cell types, including primary cultures of differentiated rat OL, Schwann cells and kidney cells, as well as neuronal and non-neuronal cell lines. All the constructs generated variable levels of chloramphenicol acetyltransferase (CAT) activity in all cell types, except in primary OL and Schwann cells, where distinct positive (PRE) and negative regulatory elements (NRE) were found to be involved in regulating cat expression. The nucleotide (nt) -53/+70 construct gave maximal activity in all cell types, except OL and Schwann cells, where sequences upstream from nt -53 were necessary for maximal promoter activity. The sequences located at nt -655 to -397 and nt -394 to -54 showed enhancer and repressor effects, respectively, in OL. In Schwann cells, sequences from -394 to -253 showed positive regulatory effects, while those between -655 to -397 and -253 to -54 showed negative regulatory effects. In the downstream region of the promoter, sequences from +20 to +70 and +70 to +200 showed strong silencer and enhancer activities specifically in OL. In gel-retardation assays using nuclear extracts prepared from several cell types and the -253 to -53 repressor sequences, specific DNA-protein complexes unique to OL were identified.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Leeds, UK.
authors.forename: A
authors.initials: A
authors.lastname: Asipu
authors.forename: G E
authors.initials: GE
authors.lastname: Blair
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D002490:Central Nervous System / Q000378:metabolism; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D017124:Conserved Sequence; D005091:Exons; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000096:biosynthesis* / Q000235:genetics; D009836:Oligodendroglia / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic; D012326:RNA Splicing; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D012583:Schwann Cells / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013552:Swine; D014407:Tumor Cells, Cultured
chemical_list: D004676:Myelin Basic Protein; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase