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Related Concept Videos

Homologous Recombination02:31

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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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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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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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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Transcript-RNA-templated DNA recombination and repair.

Havva Keskin1, Ying Shen2, Fei Huang3

  • 1School of Biology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.

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|September 5, 2014
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Endogenous transcript RNA can mediate homologous recombination for DNA double-strand break (DSB) repair in yeast. This RNA-templated repair pathway is facilitated by Rad52 and influenced by ribonucleases, impacting genomic stability.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Homologous recombination is crucial for DNA repair and genetic variation.
  • RNA molecules can participate in recombination, as observed in RNA viruses.
  • Previous studies indicated synthetic RNA can template DNA repair in cells.

Purpose of the Study:

  • To investigate if endogenous transcript RNA mediates homologous recombination with chromosomal DNA.
  • To elucidate the mechanism and factors influencing RNA-templated DNA repair.

Main Methods:

  • Development of a system to detect RNA-initiated homologous recombination in yeast Saccharomyces cerevisiae.
  • Utilizing reverse-transcription-defective yeast strains with chromosomal double-strand breaks (DSBs).
  • Assessing the role of ribonucleases H1 and H2, and the Rad52 protein.

Main Results:

  • Endogenous transcript RNA was found to mediate homologous recombination with chromosomal DNA.
  • RNA-DNA recombination was blocked by ribonucleases H1 and H2, indicating two repair pathways.
  • Proximity of transcript RNA to its DNA partner facilitated Rad52-driven recombination.
  • Yeast and human Rad52 proteins demonstrated in vitro annealing of RNA to DNA ends.

Conclusions:

  • Transcript RNA serves as a novel template for DNA double-strand break repair via homologous recombination.
  • This RNA-templated repair mechanism impacts genomic stability and plasticity.
  • The findings reveal a new layer of complexity in DNA metabolism and repair processes.