Identification of a family of cAMP response element-binding protein coactivators by genome-scale functional analysis in mammalian cells

doi:10.1073/pnas.1932773100

Identification of a family of cAMP response element-binding protein coactivators by genome-scale functional analysis in mammalian cells

^*Department of Functional Genomics, Novartis Institute for Biomedical Research, 100 Technology Square, Cambridge, MA 02139; and ^‡Genomics Institute, Novartis Research Foundation, 10675 John Jay Hopkins Drive, Suite F117, San Diego, CA 92121

Edited by Peter K. Vogt, The Scripps Research Institute, La Jolla, CA, and approved July 30, 2003 (received for review May 8, 2003)

Abstract

This report describes an unbiased method for systematically determining gene function in mammalian cells. A total of 20,704 predicted human full-length cDNAs were tested for induction of the IL-8 promoter. A number of genes, including those for cytokines, receptors, adapters, kinases, and transcription factors, were identified that induced the IL-8 promoter through known regulatory sites. Proteins that acted through a cooperative interaction between an AP-1 and an unrecognized cAMP response element (CRE)-like site were also identified. A protein, termed transducer of regulated cAMP response element-binding protein (CREB) (TORC1), was identified that activated expression through the variant CRE and consensus CRE sites. TORC1 potently induced known CREB1 target genes, bound CREB1, and activated expression through a potent transcription activation domain. A functional Drosophila TORC gene was also identified. Thus, TORCs represent a family of highly conserved CREB coactivators that may control the potency and specificity of CRE-mediated responses.

Footnotes

↵ § To whom correspondence should be addressed. E-mail: mark.labow{at}pharma.novartis.com.
↵ † V.I. and W.Z. contributed equally to this work.
This paper was submitted directly (Track II) to the PNAS office.
Abbreviations: TNF, tumor necrosis factor; CRE, cAMP response element; CRE-like, a variant CRE; CREB, cAMP response element-binding protein; TORC, transducer of regulated CREB; hTORC, human TORC, dTORC, Drosophila TORC; PKA, protein kinase A; Luc, luciferase.
Data deposition: The sequences reported in this paper have been deposited in the GenBank database [accession nos. AY360171 (hTORC1), AY360172 (hTORC2), and AY360173 (hTORC3)].