An Approach to Derive Functional Peptide Inhibitors of Transcription Factor Activity

Andrew Brennan, James T. Leech, Neil M. Kad, Jody M. Mason

Research output: Contribution to journalArticlepeer-review

Abstract

We report the development of a high-throughput, intracellular "transcription block survival" (TBS) screening platform to derive functional transcription factor antagonists. TBS is demonstrated using the oncogenic transcriptional regulator cJun, with the development of antagonists that bind cJun and prevent both dimerization and, more importantly, DNA binding remaining a primary challenge. In TBS, cognate TRE sites are introduced into the coding region of the essential gene, dihydrofolate reductase (DHFR). Introduction of cJun leads to TRE binding, preventing DHFR expression by directly blocking RNA polymerase gene transcription to abrogate cell proliferation. Peptide library screening identified a sequence that both binds cJun and antagonizes function by preventing DNA binding, as demonstrated by restored cell viability and subsequent in vitro hit validation. TBS is an entirely tag-free genotype-to-phenotype approach, selecting desirable attributes such as high solubility, target specificity, and low toxicity within a complex cellular environment. TBS facilitates rapid library screening to accelerate the identification of therapeutically valuable sequences.

Original languageEnglish
Pages (from-to)996-1006
Number of pages11
JournalJACS Au
Volume2
Issue number4
Early online date6 Apr 2022
DOIs
Publication statusPublished - 25 Apr 2022

Keywords

  • activator protein-1
  • library screening
  • peptide antagonists
  • transcription block survival
  • transcription factors

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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