A coactivator trap identifies NONO (p54nrb) as a component of the cAMP-signaling pathway
- Antonio L. Amelio*,
- Loren J. Miraglia†,
- Juliana J. Conkright‡,
- Becky A. Mercer*,
- Serge Batalov†,
- Valerie Cavett§,
- Anthony P. Orth†,
- Jennifer Busby§,
- John B. Hogenesch¶,‖, and
- Michael D. Conkright*,‖
- Departments of *Cancer Biology and
- §Molecular Therapeutics and
- ‡Translational Research Institute, The Scripps Research Institute, 5353 Parkside Drive, Jupiter, FL 33458;
- †Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121; and
- ¶Department of Pharmacology, Institute of Translational Medicine and Therapeutics, University of Pennsylvania, 3451 Walnut Street, Philadelphia, PA 19104
-
Edited by Joseph S. Takahashi, Northwestern University, Evanston, IL, and approved November 2, 2007 (received for review August 24, 2007)
Abstract
Signal transduction pathways often use a transcriptional component to mediate adaptive cellular responses. Coactivator proteins function prominently in these pathways as the conduit to the basic transcriptional machinery. Here we present a high-throughput cell-based screening strategy, termed the “coactivator trap,” to study the functional interactions of coactivators with transcription factors. We applied this strategy to the cAMP signaling pathway, which utilizes two families of coactivators, the cAMP response element binding protein (CREB) binding protein (CBP)/p300 family and the recently identified transducers of regulated CREB activity family (TORCs1–3). In addition to identifying numerous known interactions of these coactivators, this analysis identified NONO (p54nrb) as a TORC-interacting protein. RNA interference experiments demonstrate that NONO is necessary for cAMP-dependent activation of CREB target genes in vivo. Furthermore, TORC2 and NONO complex on cAMP-responsive promoters, and NONO acts as a bridge between the CREB/TORC complex and RNA polymerase II. These data demonstrate the utility of the coactivator trap by identification of a component of cAMP-mediated transcription.
Footnotes
- ‖To whom correspondence may be addressed. E-mail: hogenesc{at}mail.med.upenn.edu (for questions regarding the coactivator trap) or conkright{at}scripps.edu (for questions regarding the cAMP pathway)
-
Author contributions: A.L.A., L.J.M., J.J.C., B.A.M., S.B., A.P.O, J.B.H., and M.D.C. designed research; A.L.A., L.J.M., J.J.C., B.A.M., V.C., and M.D.C. performed research; L.J.M., J.B.H., and M.D.C. contributed new reagents/analytic tools; A.L.A., L.J.M., J.J.C., B.A.M., V.C., J.B., J.B.H., and M.D.C. analyzed data; and A.L.A., J.J.C., B.A.M., J.B.H., and M.D.C. wrote the paper.
-
The authors declare no conflict of interest.
-
This article is a PNAS Direct Submission.
-
This article contains supporting information online at www.pnas.org/cgi/content/full/0707999105/DC1.
- © 2007 by The National Academy of Sciences of the USA
