Autoregulatory sequences are revealed by complex stability screening of the mouse brn-3.0 locus.
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 approximately 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.
Department of Psychiatry, University of California San Diego, La Jolla, California 92093-0603, USA.
M
M
Trieu
J M
JM
Rhee
N
N
Fedtsova
E E
EE
Turner
D000375:Aging / Q000378:metabolism; D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D004355:Drug Stability; D004622:Embryo, Mammalian / Q000378:metabolism / Q000502:physiology; D005314:Embryonic and Fetal Development / Q000502:physiology; D005821:Genetic Techniques; D016678:Genome; D018398:Homeodomain Proteins; D006706:Homeostasis / Q000502:physiology*; D007150:Immunohistochemistry; D051379:Mice; D013997:Time Factors; D050818:Transcription Factor Brn-3; D050819:Transcription Factor Brn-3A; D050820:Transcription Factor Brn-3B; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D015533:Transcriptional Activation / Q000502:physiology
D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C494189:Pou4f1 protein, mouse; C085559:Pou4f2 protein, mouse; D050818:Transcription Factor Brn-3; D050819:Transcription Factor Brn-3A; D050820:Transcription Factor Brn-3B; D014157:Transcription Factors; D004247:DNA