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Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
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Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

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Helicase promotes replication re-initiation from an RNA transcript.

Bo Sun1,2,3, Anupam Singh4, Shemaila Sultana4

  • 1Howard Hughes Medical Institute, Cornell University, Ithaca, NY, 14853, USA. sunbo@shanghaitech.edu.cn.

Nature Communications
|June 15, 2018
PubMed
Summary
This summary is machine-generated.

T7 helicase aids DNA replication restart by disrupting stalled transcription complexes and using RNA as a primer. This reveals a new role for T7 helicase in replication re-initiation after encountering obstacles.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA replication requires overcoming obstacles like stalled transcription complexes.
  • Replication restart mechanisms after encountering obstacles are not well understood.
  • T7 replisome components, helicase and DNA polymerase, function together at the fork.

Purpose of the Study:

  • To investigate how DNA synthesis resumes after a replisome encounters an obstacle.
  • To examine the interaction between T7 helicase and non-replicating T7 DNA polymerase (DNAP).
  • To determine the role of the DNAP-helicase complex in overcoming transcription roadblocks and re-initiating replication.

Main Methods:

  • Single-molecule assays to observe molecular interactions.
  • Ensemble biochemical methods to measure enzyme activity.
  • Studying collisions between T7 replisomes and co-directional RNA polymerase (RNAP).

Main Results:

  • T7 helicase strongly interacts with a non-replicating T7 DNAP.
  • The DNAP enhances helicase processivity and unwinding rate.
  • The DNAP-helicase complex actively disrupts stalled RNAP and initiates replication using the RNA transcript as a primer.

Conclusions:

  • T7 helicase plays a novel role in replication re-initiation.
  • The DNAP-helicase partnership is crucial for overcoming transcription obstacles.
  • This mechanism facilitates the resumption of DNA synthesis downstream of impediments.