Factor-independent termination of transcription in a stretch of deoxyadenosine residues in the template DNA

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Updated: Jul 11, 2026

Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on (TRO) Approach
12:12

Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on (TRO) Approach

Published on: March 12, 2017

J Tomizawa1, H Masukata

  • 1Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.

Cell
|November 20, 1987
PubMed

Insights

Primer transcript formation in plasmid ColE1 is influenced by DNA structure. Deoxyadenosine insertions and palindromes near the replication origin affect transcription termination and RNA polymerase detachment.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Plasmid ColE1 replication relies on primer transcript synthesis.
  • Transcription can proceed in two modes: hybridized or unhybridized with template DNA.

Purpose of the Study:

  • To investigate the impact of DNA sequence modifications on transcription termination.
  • To understand the role of deoxyadenosine stretches and palindromes in regulating transcription.

Main Methods:

  • Insertion of deoxyadenosine residues into the plasmid ColE1 template DNA.
  • Analysis of primer transcript termination sites under different conditions.
  • Observing RNA polymerase detachment from the template.

Main Results:

  • A 20-deoxyadenosine insert caused >90% termination in hybridized transcripts and ~10% in unhybridized.
  • A 10-deoxyadenosine insert significantly reduced termination efficiency.
  • A preceding palindrome enhanced termination of unhybridized transcripts.
  • RNA polymerase detachment was observed upon termination of hybridized transcripts.

Conclusions:

  • DNA sequence elements, such as deoxyadenosine stretches and palindromes, significantly influence transcription termination at the ColE1 origin.
  • These structures may facilitate RNA polymerase detachment, impacting replication initiation.

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