Minimal activators that bind to the KIX domain of p300/CBP identified by phage display screening Academic Article uri icon

Overview

MeSH Major

  • Nuclear Proteins
  • Oligopeptides
  • Peptide Library
  • Trans-Activators

abstract

  • Human gene therapy approaches involving transcription factors often rely on artificial activation domains for transcriptional activation. These domains are often large (e.g., 80 amino acids for VP16), recruit multiple co-activation complexes at once, and offer no fine control over the level of transcription. In an attempt to understand the sequence and structural requirements of a minimal mammalian activator, we employed a molecular diversity approach with a peptide phage display library composed of random eight-amino acid peptides. Using the KIX domain of the mammalian co-activators p300 and CBP as target, we discovered a family of synthetic binding peptides. These peptides share significant homology with natural KIX domain ligands, and are shown to bind an overlapping, yet distinct, surface of p300/CREB-binding protein (CBP). When fused to a heterologous DNA binding domain, these synthetic peptides function as titratable, modular, and potent transcriptional activators in living cells through specific recruitment of p300/CBP, with the level of transcriptional activation proportional to the affinity of the synthetic peptide for the KIX domain. Taken together, our data demonstrate that a molecular diversity approach can be used to discover minimal, co-activator domain-specific synthetic activators, and that transcriptional activation can be modulated as desired at the level of co-activator recruitment.

publication date

  • December 2000

Research

keywords

  • Academic Article

Identity

Language

  • eng

PubMed ID

  • 11017047

Additional Document Info

start page

  • 1080

end page

  • 5

volume

  • 18

number

  • 10 SUPPL.