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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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In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
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PAR-Driven Condensation Maintains Stalled Replication Fork Stability.

Lei Zhang1, Zeyu Zhang1, Timothy R O'Leary2

  • 1Department of Pharmacology and Pharmaceutical Sciences, Alfred E. Mann School of Pharmacy and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA 90089, USA.

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Summary
This summary is machine-generated.

Poly(ADP-ribose) (PAR) forms nuclear condensates when proteasome is inhibited, co-condensing with ubiquitin chains and proteasomes. These structures protect genomic integrity by stabilizing stalled replication forks.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Genomics

Background:

  • Poly(ADP-ribose) (PAR) is a nucleic acid-like polymer involved in cellular events.
  • PAR polymerases (PARPs) catalyze PAR formation as a post-translational modification.
  • Liquid-liquid phase separation (LLPS) generates functional subcellular condensates.

Purpose of the Study:

  • To investigate the role of PAR in biomolecular condensation.
  • To identify cellular conditions and factors influencing PAR condensation.
  • To understand the functional implications of PAR condensation in stressed cells.

Main Methods:

  • Imaging-based screening of small molecules.
  • Proteasome inhibition in various cell types.
  • Co-localization studies of PAR, proteasome, and ubiquitin chains.
  • Assessment of DNA replication fork stability.

Main Results:

  • PAR undergoes LLPS upon proteasome inhibition, forming nuclear condensates.
  • These condensates involve PAR, proteasome, and ubiquitin chains, dependent on PARP2 and K6-linked ubiquitylation.
  • PAR directly interacts with ubiquitin chains for the first time.
  • Stalled DNA replication forks co-localize with these condensates.

Conclusions:

  • PAR condensation is a novel self-protective mechanism in cells under proteasomal stress.
  • PAR-proteasome-ubiquitin chain condensates stabilize stalled replication forks, maintaining genomic integrity.
  • This study provides fundamental insights into PAR condensation and its cellular functions.