Functional Organization of a Schwann Cell Enhancer

• Published in: The Journal of neuroscience Vol. 25; no. 48; pp. 11210 - 11217
• Main Authors: Denarier, Eric, Forghani, Reza, Farhadi, Hooman F, Dib, Samar, Dionne, Nancy, Friedman, Hana C, Lepage, Pierre, Hudson, Thomas J, Drouin, Regen, Peterson, Alan
• Format: Journal Article
• Language: English
• Published: United States Soc Neuroscience 30-11-2005; Society for Neuroscience
• Subjects: Amino Acid Motifs; Amino Acid Motifs - physiology; Animals; Axons; Axons - physiology; Biochemistry, Molecular Biology; Chickens; Development/Plasticity/Repair; DNA Footprinting; Enhancer Elements, Genetic; Enhancer Elements, Genetic - physiology; Humans; Hypoxanthine Phosphoribosyltransferase; Hypoxanthine Phosphoribosyltransferase - metabolism; Life Sciences; Male; Mice; Mice, Neurologic Mutants; Mice, Transgenic; Myelin Basic Protein - genetics; Myelin Basic Protein - metabolism; Myelin Basic Proteins; Neurons and Cognition; Phylogeny; Protein Structure, Tertiary; Protein Structure, Tertiary - physiology; Schwann Cells; Schwann Cells - metabolism; Signal Transduction; Signal Transduction - physiology; Species Specificity; enhancer; transgenesis; myelin basic protein; HPRT; Schwann cell; phylogenetic footprint
• ISSN: 0270-6474; 1529-2401
• DOI: 10.1523/JNEUROSCI.2596-05.2005

Myelin basic protein (MBP) gene expression is conferred in oligodendrocytes and Schwann cells by different upstream enhancers. In Schwann cells, expression is controlled by a 422 bp enhancer lying -9 kb from the gene. We show here that it contains 22 mammalian conserved motifs > or =6 bp. To investigate their functional significance, different combinations of wild-type or mutated motifs were introduced into reporter constructs that were inserted in single copy at a common hypoxanthine phosphoribosyltransferase docking site in embryonic stem cells. Lines of transgenic mice were derived, and the subsequent qualitative and quantitative expression phenotypes were compared at different stages of maturation. In the enhancer core, seven contiguous motifs cooperate to confer Schwann cell specificity while different combinations of flanking motifs engage, at different stages of Schwann cell maturation, to modulate expression level. Mutation of a Krox-20 binding site reduces the level of reporter expression, whereas mutation of a potential Sox element silences reporter expression. This potential Sox motif was also found conserved in other Schwann cell enhancers, suggesting that it contributes widely to regulatory function. These results demonstrate a close relationship between phylogenetic footprints and regulatory function and suggest a general model of enhancer organization. Finally, this investigation demonstrates that in vivo functional analysis, supported by controlled transgenesis, can be a robust complement to molecular and bioinformatics approaches to regulatory mechanisms.