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J Biol Chem, Vol. 273, Issue 35, 22241-22247, August 28, 1998

Association of Pur with RNAs Homologous to 7 SL Determines Its Binding Ability to the Myelin Basic Protein Promoter DNA Sequence

Anna Tretiakova, Gary L. Gallia, Natalia Shcherbik, Bradford Jameson, Edward M. Johnson^§, Shohreh Amini, and Kamel Khalili

From the  Center for NeuroVirology and NeuroOncology, Allegheny University of the Health Sciences, Philadelphia, Pennsylvania 19102 and the ^§ Department of Pathology, Mount Sinai School of Medicine, New York, New York 10029

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 Pur, 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 Pur. Results from band shift competition studies indicate that Pur-associated RNA (PU-RNA), inhibits the interaction of immunopurified Pur 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 Pur 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 Pur that are important for Pur/PU-RNA association. Production of Pur at the early stage of brain development and its association with PU-RNA at this stage, when Pur exhibits poor binding ability to the MB1 DNA sequence, suggests that PU-RNA may function as a co-factor that negatively regulates Pur interaction with the MBP promoter sequence.

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