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The DNA Replication Fork01:02

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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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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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The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
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A replication stress safeguard provided by the Elg1 Replication Factor C-like complex.

Pallavi Bose1, Soumitra Sau1

  • 1Amity Institute of Biotechnology, Amity University Kolkata, West Bengal 700135, India.

Genetics
|September 23, 2025
PubMed
Summary

The Elg1 Replication Factor C-like complex (Elg1-RLC) prevents replication fork collapse during DNA damage. It unloads proliferating cell nuclear antigen (PCNA) to maintain genome stability under replication stress.

Keywords:
9–1–1/Rad24DDR sensors/mediatorsDNA damage response (DDR)Elg1MMSMec1/Rad53PCNAreplication forksreplication stress

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • The Elg1 Replication Factor C-like complex (Elg1-RLC) unloads proliferating cell nuclear antigen (PCNA).
  • Elg1-RLC is implicated in DNA replication and repair across species.
  • PCNA unloading is crucial for managing DNA damage during replication.

Purpose of the Study:

  • To investigate the role of Elg1-RLC's PCNA unloading activity in response to DNA alkylating agents.
  • To elucidate the mechanism by which Elg1-RLC maintains replication fork (RF) integrity under stress.
  • To determine if Elg1-RLC functions through canonical or noncanonical S-phase checkpoint pathways.

Main Methods:

  • Utilized disassembly-prone PCNA mutants to study Elg1-RLC function.
  • Assessed cell viability of elg1Δ-DDR double mutants exposed to methyl-methanesulfonate (MMS).
  • Analyzed Rad52 foci formation and effects of deoxynucleoside triphosphate (dNTP) levels on mutant rescue.

Main Results:

  • Elg1-RLC's PCNA unloading counters MMS-induced replication fork slowing.
  • elg1Δ-DDR mutants exhibit sensitivity to MMS, indicative of replication fork collapse.
  • Elevating dNTP levels failed to rescue elg1Δ-DDR mutants, ruling out dNTP shortage as the cause of fork collapse.
  • Rad52 foci accumulation in elg1Δ-DDR cells supports the occurrence of replication fork collapse.

Conclusions:

  • Elg1-RLC plays a critical role in preventing replication fork collapse under replication stress.
  • Elg1-RLC likely regulates the S-phase checkpoint via a noncanonical pathway.
  • Timely PCNA unloading by Elg1-RLC coordinates DNA damage response pathways to safeguard genome integrity.