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Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets
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Identification of a family of cAMP response element-binding protein coactivators by genome-scale functional analysis

Vadim Iourgenko1, Wenjun Zhang, Craig Mickanin

  • 1Department of Functional Genomics, Novartis Institute for Biomedical Research, 100 Technology Square, Cambridge, MA 02139, USA.

Proceedings of the National Academy of Sciences of the United States of America
|September 25, 2003
PubMed

Insights

Researchers developed a method to discover gene functions in mammalian cells by testing human cDNAs for IL-8 promoter induction. They identified novel proteins, including TORC1, a conserved CREB coactivator crucial for gene regulation.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Determining gene function is crucial for understanding mammalian cell biology.
  • Systematic methods are needed to identify genes regulating specific cellular responses.

Purpose of the Study:

  • To develop an unbiased method for systematically determining gene function in mammalian cells.
  • To identify genes and proteins that regulate the Interleukin-8 (IL-8) promoter.

Main Methods:

  • Screened 20,704 predicted human full-length cDNAs for IL-8 promoter induction.
  • Analyzed identified genes for roles as cytokines, receptors, adapters, kinases, and transcription factors.
  • Investigated cooperative interactions between transcription factors and identified novel coactivators.

Main Results:

  • Identified multiple genes inducing the IL-8 promoter via known regulatory sites.
  • Discovered proteins mediating cooperative interactions with AP-1 and a CRE-like site.
  • Identified transducer of regulated cAMP response element-binding protein (TORC1) as a potent CREB coactivator.
  • Demonstrated TORC1's ability to activate CREB1 target genes and bind CREB1.

Conclusions:

  • TORC1 is a conserved CREB coactivator family that enhances CREB-mediated gene responses.
  • This systematic approach is effective for discovering gene functions and regulatory pathways.
  • TORC1 plays a significant role in controlling the potency and specificity of cAMP response element (CRE)-mediated transcription.

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