Polymerase Arrest at the λP R Promoter during Transcription Initiation*

During transcription initiation byEscherichia coli RNA polymerase, a fraction of the homogeneous enzyme population has been kinetically shown to form two types of nonproductive complexes at some promoters: moribund complexes, which produce only abortive transcripts, and fully inactive ternary complexes (Kubori, T., and Shimamoto, N. (1996) J. Mol. Biol. 256, 449–457). Here we report biochemical isolation of the complexes arrested at the λP R promoter and an analysis of their structure by DNA and protein footprintings. We found that the isolated promoter-arrested complexes retain a stoichiometric amount of ς70 subunit. Exonuclease III footprints of the arrested complexes are backtracked compared with that of the binary complex, and KMnO4 footprinting reveals a decrease in the melting of DNA in the promoter region. Protein footprints of the retained ς70 have shown a more exposed conformation in region 3, compared with binary complexes. This feature is similar to that of the complexes arrested in inactive state during transcription elongation, indicating the existence of a common inactivating mechanism during transcription initiation and elongation. The possible involvement of the promoter arrest in transcriptional regulation is discussed.

• Publication year

  2000

• Venue

  Journal of Biological Chemistry

• Publication date

  2000-04-14

• Fields of study

  Biology

• Identifiers
  DOI 10.1074/JBC.275.15.10899
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar

• No claims are published for this paper.

• No concepts are published for this paper.

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