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One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
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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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RAD18 methylation by the methyltransferase SETD6 attenuates DNA breaks.

Lital Estrella Weil1, Michal Feldman1, Jennifer Van Duine2

  • 1The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, P.O.B. 653, Be'er-Sheva, 84105, Israel.

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This summary is machine-generated.

The SETD6 methyltransferase interacts with and methylates RAD18, a DNA repair protein. This methylation regulates RAD18

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • DNA Repair Mechanisms

Background:

  • SETD6 is a SET-domain-containing methyltransferase involved in post-translational modifications.
  • RAD18 is a crucial protein in the DNA damage repair pathway.
  • Protein methylation regulates protein function, stability, and interactions.

Purpose of the Study:

  • To investigate the interaction between SETD6 and RAD18.
  • To determine if SETD6 methylates RAD18 and elucidate the functional consequences of this modification.
  • To understand the role of SETD6-mediated RAD18 methylation in maintaining genomic integrity.

Main Methods:

  • Protein microarray technology to identify interactors.
  • ELISA and immunoprecipitation assays to confirm interaction and methylation.
  • Mass spectrometry and site-directed mutagenesis to identify methylation sites.
  • Analysis of DNA damage markers (γH2AX) and comet assays in SETD6 knockout cells.

Main Results:

  • RAD18 was identified as a direct interactor and substrate of SETD6.
  • RAD18 undergoes mono-methylation at K73 and K406 residues by SETD6.
  • SETD6-mediated methylation influences RAD18's nuclear localization.
  • SETD6 depletion leads to increased DNA damage markers and DNA breaks.

Conclusions:

  • SETD6-mediated methylation of RAD18 is essential for attenuating DNA breaks.
  • This methylation regulates RAD18's cellular localization and function.
  • SETD6 plays a critical role in maintaining genomic integrity through RAD18 regulation.