Design of a Temperature-Responsive Transcription Terminator

ACS Synth Biol. 2018 Feb 16;7(2):613-621. doi: 10.1021/acssynbio.7b00356. Epub 2017 Dec 12.

Abstract

RNA structures regulate various steps in gene expression. Transcription in bacteria is typically terminated by stable hairpin structures. Translation initiation can be modulated by metabolite- or temperature-sensitive RNA structures, called riboswitches or RNA thermometers (RNATs), respectively. RNATs control translation initiation by occlusion of the ribosome binding site at low temperatures. Increasing temperatures destabilize the RNA structure and facilitate ribosome access. In this study, we exploited temperature-responsive RNAT structures to design regulatory elements that control transcription termination instead of translation initiation in Escherichia coli. In order to mimic the structure of factor-independent intrinsic terminators, naturally occurring RNAT hairpins were genetically engineered to be followed by a U-stretch. Functional temperature-responsive terminators (thermoterms) prevented mRNA synthesis at low temperatures but resumed transcription after a temperature upshift. The successful design of temperature-controlled terminators highlights the potential of RNA structures as versatile gene expression control elements.

Keywords: RNA thermometer; gene expression; regulatory RNA; synthetic biology; temperature; transcription termination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Escherichia coli* / chemistry
  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Gene Expression Regulation, Bacterial*
  • Hot Temperature*
  • RNA Folding*
  • RNA, Bacterial* / biosynthesis
  • RNA, Bacterial* / chemistry
  • RNA, Bacterial* / genetics
  • Terminator Regions, Genetic*

Substances

  • RNA, Bacterial