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Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

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For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
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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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The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
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Related Experiment Video

Updated: Dec 28, 2025

Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence
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Physiological Tolerance to ssDNA Enables Strand Uncoupling during DNA Replication.

Amaia Ercilla1, Jan Benada1, Sampath Amitash1

  • 1Center for Chromosome Stability, Institute for Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen 2200, Denmark.

Cell Reports
|February 21, 2020
PubMed
Summary

Human DNA polymerases can work independently on DNA strands, allowing for uncoupled replication. This process, while robust, creates a vulnerability that can be exploited for cancer therapy.

Keywords:
ATRCD437DNA replicationPOLA1RPAlagging strandpolymerase alphareplication catastrophessDNAstrand uncoupling

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • The dogma of coordinated leading and lagging strand synthesis during DNA replication has been challenged by in vitro studies.
  • Previous understanding assumed coordinated synthesis prevents DNA strand uncoupling and single-stranded DNA (ssDNA) accumulation.

Purpose of the Study:

  • To investigate whether human DNA polymerases can function independently in vivo.
  • To determine the physiological implications and cellular tolerance to ssDNA during uncoupled DNA replication.

Main Methods:

  • Inhibition of POLA1 (a key enzyme in DNA replication) in human cells.
  • Analysis of ssDNA accumulation and cellular responses, including stress responses and RPA (Replication Protein A) levels.

Main Results:

  • Human DNA polymerases can function independently in vivo, leading to strand uncoupling.
  • Cells tolerate significant ssDNA accumulation at the lagging strand when POLA1 is inhibited, without immediate stress response.
  • Limited POLA1 activity supports DNA duplication with exacerbated uncoupling, provided RPA is sufficient.

Conclusions:

  • DNA replication can occur in an uncoupled manner in human cells, supported by cellular ssDNA tolerance.
  • This uncoupled replication, while physiologically supported, presents a vulnerability that can be therapeutically targeted in cancer treatment.