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CREB-binding protein activates transcription through multiple domains

D L Swope1, C L Mueller, J C Chrivia

  • 1Department of Pharmacological and Physiological Sciences, Saint Louis University School of Medicine, St. Louis, Missouri 63104, USA.

The Journal of Biological Chemistry
|November 8, 1996
PubMed
Summary

The N-terminal half of CREB-binding protein (CBP) contains a transcriptional activation domain (TAD) sufficient for CREB-mediated transcription. This domain binds TBP and its activity is cell-type specific in response to PKA.

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Protein Function

Background:

  • CREB-binding protein (CBP) is a coactivator for various transcription factors.
  • The specific regions of CBP responsible for transcriptional activation were previously unidentified.

Purpose of the Study:

  • To identify and characterize the transcriptional activation domains (TADs) within CBP.
  • To investigate the interaction of CBP TADs with TATA-binding protein (TBP).
  • To determine the cell-type specificity of PKA-mediated regulation of CBP TADs.

Main Methods:

  • Deletion analysis to identify functional domains within CBP.
  • Squelching assays to assess the role of TADs in transcription.
  • In vitro binding assays to examine CBP TAD-TBP interactions.

Related Experiment Videos

  • GAL-CBP chimera constructs to study C-terminal TADs.
  • Reporter gene assays in various cell lines (PC-12, F9, COS-7) to assess transcriptional activity.

    • Main Results:

    • The N-terminal half of CBP possesses a potent TAD sufficient for CREB-mediated transcription.
    • This N-terminal TAD directly binds TBP.
    • Two distinct C-terminal TADs were identified adjacent to the c-Fos binding site.
    • PKA enhances the transcriptional activity of the N-terminal TADs in PC-12 cells, but not in F9 or COS-7 cells, indicating cell-type specific regulation.

    Conclusions:

    • CBP contains multiple TADs, with the N-terminal region being crucial for CREB-mediated transcription and TBP binding.
    • The regulation of CBP activity by PKA is cell-type specific.
    • These findings elucidate the functional organization of CBP's transcriptional activation capabilities.