Cloning of a novel Olf-1/EBF-like gene, O/E-4, by degenerate oligo-based direct selection.
The Olf-1/EBF (O/E) family of transcription factors plays important roles in neural development and B-cell maturation. We have utilized a variation of the direct selection scheme (Degenerate Oligo-based Direct Selection, or DODS) to identify a fourth member of the O/E family (O/E-4). We demonstrated that, similar to previously identified mouse O/E proteins, O/E-4 is expressed in the neuronal and basal cell layers of mouse olfactory epithelium. However, unlike other O/E members, O/E-4 expression is absent in the vomeronasal organ. O/E-4 can partner with the other O/Es to form heterodimers that are capable of binding an Olf-1 consensus site in vitro. In contrast to other O/E family members, the O/E-4 protein only weakly activates transcription in a reporter assay. In summary, we have identified O/E-4 utilizing a novel homology screening method. Our results suggest that O/E-4 may interact with other O/E family members to regulate gene expression in the olfactory sensory neurons.
Department of Molecular Biology and Genetics, The Howard Hughes Medical Institute, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Song S
SS
Wang
Alexander G
AG
Betz
Randall R
RR
Reed
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics*; D015723:Gene Library; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009831:Olfactory Mucosa / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism*; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics*; D019147:Vomeronasal Organ / Q000378:metabolism
D004268:DNA-Binding Proteins; C081976:EBF1 protein, human; C489921:Ebf1 protein, mouse; D009841:Oligonucleotides; D015534:Trans-Activators