Regulation of mouse myelin basic protein gene transcription by a sequence-specific single-stranded DNA-binding protein in vitro.
Cell- and stage-specific transcription of the myelin basic protein (MBP)-encoding gene (Mbp) in brain is regulated by arrays of cis-acting regulatory elements positioned at the 5'-flanking region of the gene. The proximal element between nucleotides -14 to -50, termed MB1, has previously been shown to have an important role in the cell-type-specific transcription of Mbp in cells derived from the central nervous system (CNS). Here, we utilized band-shift and in vitro transcription assays to examine the ability of MEF-2, an expression factor encoded by a cDNA isolated from mouse brain, in binding to the MB1 element and regulating transcription of the Mbp promoter. Results from the band-shift assays indicated that the bacterially produced MEF-2 recognizes specific nt in the MB1 motif, and its binding to these nt reduces the overall transcriptional activity of Mbp in a cell-free extract.
Steplewski
A
A
Jefferson Institute of Molecular Medicine, Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Haas
S
S
Amini
S
S
Khalili
K
K
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D002474:Cell-Free System; D004268:DNA-Binding Proteins / Q000378:metabolism*; D064326:MEF2 Transcription Factors; D051379:Mice; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000096:biosynthesis* / Q000235:genetics*; D018006:Myogenic Regulatory Factors; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D012097:Repressor Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D064326:MEF2 Transcription Factors; D004676:Myelin Basic Protein; D018006:Myogenic Regulatory Factors; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors