Alternative transcription cycle for bacterial RNA polymerase

Nat Commun. 2020 Jan 23;11(1):448. doi: 10.1038/s41467-019-14208-9.

Abstract

RNA polymerases (RNAPs) transcribe genes through a cycle of recruitment to promoter DNA, initiation, elongation, and termination. After termination, RNAP is thought to initiate the next round of transcription by detaching from DNA and rebinding a new promoter. Here we use single-molecule fluorescence microscopy to observe individual RNAP molecules after transcript release at a terminator. Following termination, RNAP almost always remains bound to DNA and sometimes exhibits one-dimensional sliding over thousands of basepairs. Unexpectedly, the DNA-bound RNAP often restarts transcription, usually in reverse direction, thus producing an antisense transcript. Furthermore, we report evidence of this secondary initiation in live cells, using genome-wide RNA sequencing. These findings reveal an alternative transcription cycle that allows RNAP to reinitiate without dissociating from DNA, which is likely to have important implications for gene regulation.

Publication types

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

MeSH terms

  • Adenosine Triphosphate / genetics
  • Cytidine Triphosphate / genetics
  • DNA / genetics
  • DNA / metabolism
  • DNA, Antisense / genetics
  • DNA-Directed RNA Polymerases / chemistry
  • DNA-Directed RNA Polymerases / genetics*
  • DNA-Directed RNA Polymerases / metabolism
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Microscopy, Fluorescence
  • Promoter Regions, Genetic
  • Single Molecule Imaging
  • Transcription, Genetic*

Substances

  • DNA, Antisense
  • Cytidine Triphosphate
  • Adenosine Triphosphate
  • DNA
  • DNA-Directed RNA Polymerases