Association of Puralpha with RNAs homologous to 7 SL determines its binding ability to the myelin basic protein promoter DNA sequence.
Cell type and developmental stage expression of the myelin basic protein (MBP) gene in mouse brain is regulated at the transcriptional level. Earlier studies from our laboratory have led to the identification of a DNA binding protein from mouse brain, named Puralpha, which interacts with the MB1 regulatory motif of the MBP and stimulates its transcription in glial cells. In this report, we demonstrate that a cellular RNA, with significant homology to 7 SL RNA is associated with Puralpha. Results from band shift competition studies indicate that Puralpha-associated RNA (PU-RNA), inhibits the interaction of immunopurified Puralpha with the MB1 DNA sequence. Results from Northern blot studies indicated that PU-RNA is expressed during various stages of brain development. Of interest, this RNA was found in association with Puralpha that was produced in the mouse brain at the early stage of brain development. Results from Northwestern analysis using a PU-RNA probe identified the regions within Puralpha that are important for Puralpha/PU-RNA association. Production of Puralpha at the early stage of brain development and its association with PU-RNA at this stage, when Puralpha exhibits poor binding ability to the MB1 DNA sequence, suggests that PU-RNA may function as a co-factor that negatively regulates Puralpha interaction with the MBP promoter sequence.
Center for NeuroVirology and NeuroOncology, Allegheny University of the Health Sciences, Philadelphia, Pennsylvania 19102, USA.
A
A
Tretiakova
G L
GL
Gallia
N
N
Shcherbik
B
B
Jameson
E M
EM
Johnson
S
S
Amini
K
K
Khalili
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D001921:Brain / Q000254:growth & development / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein; D004247:DNA; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000235:genetics*; D009419:Nerve Tissue Proteins; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors
D017362:Cyclic AMP Response Element-Binding Protein; D004268:DNA-Binding Proteins; D004676:Myelin Basic Protein; D009419:Nerve Tissue Proteins; C121485:Pura protein, mouse; D016601:RNA-Binding Proteins; D014157:Transcription Factors; D012313:RNA; D004247:DNA