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Nucleotide Excision Repair01:08

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Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
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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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In order to be passed through generations, genomic DNA must be undamaged and error-free. However, every day, DNA in a cell undergoes several thousand to a million damaging events by natural causes and external factors. Ionizing radiation such as UV rays, free radicals produced during cellular respiration, and hydrolytic damage from metabolic reactions can alter the structure of DNA. Damages caused include single-base alteration, base dimerization, chain breaks, and cross-linkage.
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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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Investigation of Protein Recruitment to DNA Lesions Using 405 Nm Laser Micro-irradiation
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An extended sequence specificity for UV-induced DNA damage.

Long H Chung1, Vincent Murray1

  • 1School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia.

Journal of Photochemistry and Photobiology. B, Biology
|November 18, 2017
PubMed
Summary
This summary is machine-generated.

This study precisely mapped UV-induced DNA damage sequence specificity to a hexanucleotide level. Neighboring nucleotides, particularly A and C, significantly influence UV DNA damage occurrence.

Keywords:
Consensus sequenceCyclopyrimidine dimerDNA sequence specificityEnd-labellingLinear amplificationUV photoproducts

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Area of Science:

  • Molecular Biology
  • Photochemistry
  • Genetics

Background:

  • UV radiation causes DNA damage, forming cyclobutane pyrimidine dimers (CPDs) and pyrimidine(6-4)pyrimidone photoproducts (6-4PPs).
  • Understanding UV DNA damage sequence specificity is crucial for DNA repair mechanisms and photoprotection strategies.

Purpose of the Study:

  • To determine UV-induced DNA damage sequence specificity with enhanced precision and accuracy.
  • To extend the analysis of UV sequence specificity from dinucleotide/trinucleotide to hexanucleotide levels.
  • To investigate the influence of neighboring nucleotides on UV DNA damage.

Main Methods:

  • Utilized capillary electrophoresis with laser-induced fluorescence.
  • Employed end-labeling (for 6-4PPs) and polymerase stop/linear amplification assays.
  • Analyzed UV damage in a random sequence mitochondrial DNA target.

Main Results:

  • Identified consensus sequences for UV damage: 5'-GCTC*AC (end-labeling) and 5'-TCTT*AC (linear amplification).
  • Determined highest dinucleotide frequencies for UV damage: 5'-TC*, 5'-TT*, and 5'-CC* (end-labeling); 5'-TT* (linear amplification).
  • Found that an 'A' at position 1 and 'C' at position 2 enhance UV-induced DNA damage.

Conclusions:

  • UV-induced DNA damage exhibits sequence specificity extending to the hexanucleotide level.
  • Neighboring nucleotides play a significant role in modulating the extent of UV DNA damage.
  • The findings provide a more detailed understanding of UV mutagenesis and DNA repair pathways.