Structural basis of bacterial σ28-mediated transcription reveals roles of the RNA polymerase zinc-binding domain

Wei Shi, Wei Zhou, Baoyue Zhang, Shaojia Huang, Yanan Jiang, Abigail Schammel, Yangbo Hu, Bin Liu

Research output: Contribution to journalArticlepeer-review

Abstract

In bacteria, σ28 is the flagella-specific sigma factor that targets RNA polymerase (RNAP) to control the expression of flagella-related genes involving bacterial motility and chemotaxis. However, the structural mechanism of σ28-dependent promoter recognition remains uncharacterized. Here, we report cryo-EM structures of E. coli σ28-dependent transcribing complexes on a complete flagella-specific promoter. These structures reveal how σ28-RNAP recognizes promoter DNA through strong interactions with the −10 element, but weak contacts with the −35 element, to initiate transcription. In addition, we observed a distinct architecture in which the β′ zinc-binding domain (ZBD) of RNAP stretches out from its canonical position to interact with the upstream non-template strand. Further in vitro and in vivo assays demonstrate that this interaction has the overall effect of facilitating closed-to-open isomerization of the RNAP–promoter complex by compensating for the weak interaction between σ4 and −35 element. This suggests that ZBD relocation may be a general mechanism employed by σ70 family factors to enhance transcription from promoters with weak σ4/−35 element interactions.

Original languageEnglish (US)
Article numbere104389
JournalEMBO Journal
Volume39
Issue number14
DOIs
StatePublished - Jul 15 2020

Keywords

  • Cryo-EM
  • flagellar gene regulation
  • transcription initiation complex
  • ZBD relocation
  • σ

PubMed: MeSH publication types

  • Journal Article

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