MyEF-3, a developmentally controlled brain-derived nuclear protein which specifically interacts with myelin basic protein proximal regulatory sequences

Abstract

Myelin basic protein (MBP) gene contains the upstream regulatory sequence that confers cell type- and stage-specific transcription to MBP expression in oligodendrocytes during brain development. The MB1 regulatory motif located between -14 to -50 with respect to transcription start site binds to a brain derived nuclear protein and plays an important role in transcriptional activation of the MBP promoter in transfection assay. Here, we report the isolation of a recombinant cDNA clone, termed myelin expression factor-3, (MyEF-3) from a mouse brain expression library that encodes a novel protein which interacts with the MBP MB1 domain. Computer assisted evaluations of the MyEF-3 sequence revealed several interesting features including four sites for phosphorylation by casein kinase II, a transmembrane domain at the N-terminus, a nuclear localization signal and a Zinc finger domain at the carboxyl terminal. Results from Western and band shift assays indicate that MyEF-3 binds efficiently to double-stranded MB1 as well as the single-stranded non-coding strand of MB1. The use of short DNA fragments encompassing the nucleotide base substitutions across the MB1 domain in competition band shift assay revealed that the ten nucleotide sequence, 5'-GCCTGTCTTT-3' is important for binding of MyEF-3 to DNA. Results from Northern blot studies demonstrate that expression of MyEF-3 is restricted to brain and developmentally regulated during brain maturation. The biological importance of MyEF-3 in the cell type- and stage-specific expression of MBP during brain development is discussed.