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DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
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An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication...
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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
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DNA Polymerase Locks Replication Fork Under Stress.

Xiaomeng Jia1,2, James T Inman1,2, Anupam Singh3

  • 1Howard Hughes Medical Institute, Cornell University, Ithaca, NY 14853, USA.

Biorxiv : the Preprint Server for Biology
|October 17, 2024
PubMed
Summary
This summary is machine-generated.

DNA polymerase (DNAP) inactivates DNA replication when working against replication fork stress. DNAP

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • DNA replication requires unwinding of parental DNA by the replisome.
  • DNA polymerase (DNAP) and helicase are key proteins in the replisome.
  • The consequences of DNAP functioning independently of helicase are not fully understood.

Purpose of the Study:

  • To investigate the effects of DNAP operating under replication fork stress.
  • To understand the interaction between DNAP and the replication fork.
  • To elucidate mechanisms protecting replication forks during stress.

Main Methods:

  • Examined DNAP behavior under simulated replication fork stress.
  • Analyzed DNAP interactions with leading and lagging DNA strands.
  • Investigated the role of DNAP exonuclease activity in fork stability.

Main Results:

  • Prolonged DNAP exposure to fork stress leads to replication inactivation.
  • DNAP strongly interacts with DNA strands, causing replication fork locking.
  • DNAP's reverse translocation (exonuclease activity) is crucial for preventing fork inactivation.
  • Helicase addition cannot reverse the observed fork locking.

Conclusions:

  • DNAP interaction with the replication fork under stress can lead to fork locking and inactivation.
  • DNAP exonuclease activity is essential for maintaining replication fork activity during stress.
  • Understanding DNAP-fork dynamics is key to comprehending replication stress responses.