Visualizing translocation dynamics and nascent transcript errors in paused RNA polymerases in vivo.

ABSTRACT:

Background

Transcription elongation is frequently interrupted by pausing signals in DNA, with downstream effects on gene expression. Transcription errors also induce prolonged pausing, which can lead to a destabilized genome by interfering with DNA replication. Mechanisms of pausing associated with translocation blocks and misincorporation have been characterized in vitro, but not in vivo.

Results

We investigate the pausing pattern of RNA polymerase (RNAP) in Escherichia coli by a novel approach, combining native elongating transcript sequencing (NET-seq) with RNase footprinting of the transcripts (RNET-seq). We reveal that the G-dC base pair at the 5' end of the RNA-DNA hybrid interferes with RNAP translocation. The distance between the 5' G-dC base pair and the 3' end of R

SUBMITTER: Imashimizu M 

PROVIDER: S-EPMC4457086 | biostudies-literature | 2015 May

REPOSITORIES: biostudies-literature