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The Journal of Neuroscience, August 1, 1999, 19(15):6549-6558

Autoregulatory Sequences are Revealed by Complex Stability Screening of the Mouse brn-3.0 Locus

May Trieu^{1, 2}, Jerry M. Rhee¹, Natalia Fedtsova¹, and Eric E. Turner^{1, 2}

¹ Department of Psychiatry, University of California San Diego, La Jolla, California 92093-0603 and ² San Diego Veterans Affairs Medical Center, San Diego, California 92121

The POU-IV or Brn-3 class of transcription factors exhibit conserved structure, DNA-binding properties, and expression in specific subclasses of neurons across widely diverged species. In the mouse CNS, Brn-3.0 expression characterizes specific neurons from neurogenesis through the life of the cell. This irreversible activation of expression suggests positive autoregulation. To search for cis-acting elements that could mediate autoregulation we used a novel method, complex stability screening, which we applied to rapidly identify functional Brn-3.0 recognition sites within a large genomic region encompassing the mouse brn-3.0 locus. This method is based on the observation that the kinetic stability of Brn-3.0 complexes with specific DNA sequences, as measured by their dissociation half-lives, is highly correlated with the ability of those sequences to mediate transcriptional activation by Brn-3.0. The principal Brn-3.0 autoregulatory region lies ~5 kb upstream from the Brn-3.0 transcription start site and contains multiple Brn-3.0-binding sites that strongly resemble the optimal binding site for this protein class. This region also mediates transactivation by the closely related protein Brn-3.2, suggesting a regulatory cascade of POU proteins in specific neurons in which Brn-3.2 expression precedes Brn-3.0.

Key words: Brn-3; POU-domain; autoregulation; transcription factor; homeodomain; dissociation kinetics; retina; habenula; inferior olive; tectum

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Copyright © 1999 Society for Neuroscience  0270-6474/99/19156549-10$05.00/0

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