An RNA-based transcription activator derived from an inhibitory aptamer

Nucleic Acids Res. 2010 Apr;38(7):2378-86. doi: 10.1093/nar/gkp1227. Epub 2010 Jan 12.

Abstract

According to the recruitment model of transcriptional activation, an activator helps initiate transcription by bringing the RNA polymerase to a specific location on the DNA through interaction with components of the transcriptional machinery. However, it is difficult to isolate and define the activities of specific activator-target pairs experimentally through rearranging existing protein parts. Here we designed and constructed an RNA-based transcriptional activator to study specificity from both sides of the activator-target interface. Utilizing a well-characterized site-specific RNA aptamer for TFIIB, we were able to delineate some key features of this process. By rationally converting an inhibitory aptamer into the activation domain of the activator, we also introduced a new source of submolecular building blocks to synthetic biology.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aptamers, Nucleotide / chemistry*
  • Aptamers, Nucleotide / genetics
  • Aptamers, Nucleotide / metabolism
  • Base Sequence
  • Molecular Sequence Data
  • RNA / chemistry*
  • Saccharomyces cerevisiae / genetics
  • Trans-Activators / chemistry*
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • Transcription Factor TFIIB / metabolism
  • Transcriptional Activation*

Substances

  • Aptamers, Nucleotide
  • Trans-Activators
  • Transcription Factor TFIIB
  • RNA