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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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Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

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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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DNA Damage can Stall the Cell Cycle02:36

DNA Damage can Stall the Cell Cycle

10.4K
In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
10.4K
DNA Damage Can Stall the Cell Cycle02:36

DNA Damage Can Stall the Cell Cycle

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In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
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Homologous Recombination02:31

Homologous Recombination

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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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S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

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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).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of...
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Related Experiment Video

Updated: Apr 19, 2026

Visualization of DNA Repair Proteins Interaction by Immunofluorescence
07:55

Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Published on: June 26, 2020

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CSB interacts with SNM1A and promotes DNA interstrand crosslink processing.

Teruaki Iyama1, Sook Y Lee2, Brian R Berquist3

  • 1Laboratory of Molecular Gerontology, National Institute on Aging, Intramural Research Program, National Institutes of Health, Baltimore, MD 21224, USA.

Nucleic Acids Research
|December 16, 2014
PubMed
Summary
This summary is machine-generated.

Cockayne syndrome involves premature aging and degeneration. Researchers found that CSB protein coordinates DNA repair with SNM1A, and defects in this process may cause CS pathologies.

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Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins
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Visualization of DNA Repair Proteins Interaction by Immunofluorescence
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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
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Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins
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Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Cockayne syndrome (CS) is a premature aging disorder with multisystem degeneration.
  • CS is classified into complementation groups A and B.
  • The CSB protein's function in DNA repair is not fully understood.

Purpose of the Study:

  • To investigate the interaction between CSB and SNM1A.
  • To elucidate the role of CSB and SNM1A in DNA repair mechanisms.
  • To understand the contribution of DNA repair defects to Cockayne syndrome pathologies.

Main Methods:

  • Yeast two-hybrid screening to identify interacting proteins.
  • Recombinant protein interaction assays and in vitro exonuclease activity tests.
  • Fluorescent protein tagging, confocal microscopy, and laser microirradiation for in vivo studies.
  • Comet assay and γ-H2AX foci analysis to assess DNA repair efficiency.

Main Results:

  • SNM1A, a 5'-3' exonuclease, directly interacts with CSB.
  • CSB modulates SNM1A exonuclease activity and forms a complex with it in human cells.
  • Both CSB and SNM1A are recruited to DNA interstrand crosslink (ICL) damage sites, dependent on transcription.
  • SNM1A recruitment is impaired in CSB-deficient cells.
  • CSB-deficient neural cells show increased sensitivity to DNA crosslinking agents and delayed ICL repair.

Conclusions:

  • CSB coordinates the resolution of ICLs, potentially via a transcription-associated repair pathway involving SNM1A.
  • Defects in CSB-SNM1A-mediated DNA repair may contribute to the degenerative pathologies observed in Cockayne syndrome, particularly in non-cycling cells.
  • This study reveals a novel role for SNM1A in conjunction with CSB in DNA repair and provides insights into CS pathogenesis.