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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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Separation of Single-stranded DNA, Double-stranded DNA and RNA from an Environmental Viral Community Using Hydroxyapatite Chromatography
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Constitutively active Artemis nuclease recognizes structures containing single-stranded DNA configurations.

Nicholas R Pannunzio1, Michael R Lieber2

  • 1Department of Pathology, Keck School of Medicine of University of Southern California, Los Angeles, CA, 90089, USA; Norris Comprehensive Cancer Center, Keck School of Medicine of University of Southern California, Los Angeles, CA, 90089, USA.

DNA Repair
|August 5, 2019
PubMed
Summary
This summary is machine-generated.

The constitutively active Artemis-413 enzyme detects DNA structures under torsional stress. Its activity reveals distinct cellular responses to topoisomerase mutations versus drug-induced inhibition, highlighting its utility in DNA repair studies.

Keywords:
ArtemisDNA double-strand breaksDNA repairRecombinationSaccharomyces cerevisiaeTopoisomerases

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Artemis nuclease cleaves single-stranded to double-stranded DNA (ss/dsDNA) junctions.
  • It plays a crucial role in DNA double-strand break (DSB) repair via non-homologous end joining (NHEJ).
  • A constitutively active truncated form, Artemis-413, is available for in vitro and in vivo studies.

Purpose of the Study:

  • To utilize Artemis-413 to detect ss/dsDNA structures formed under topological stress.
  • To investigate the cellular consequences of altered topoisomerase activity on DNA structure and repair.
  • To differentiate the cellular effects of genetic topoisomerase ablation versus pharmacological inhibition.

Main Methods:

  • Overexpression of Artemis-413 in yeast strains with impaired topoisomerase activity.
  • Treatment of cells with camptothecin, a topoisomerase I inhibitor.
  • Quantification of DNA double-strand breaks (DSBs) in response to genetic and chemical perturbations.

Main Results:

  • Yeast cells overexpressing Artemis-413 with mutated topoisomerases showed increased DSBs, suggesting increased ss/dsDNA structures.
  • Camptothecin treatment led to increased DSBs, but co-expression of Artemis-413 reduced these DSBs.
  • Observed contrasting outcomes highlight distinct cellular responses to topoisomerase dysfunction.

Conclusions:

  • Increased torsional stress leads to DNA structures recognized by Artemis, potentially causing DSBs.
  • Topoisomerase genetic mutations and camptothecin-induced inhibition elicit different cellular responses.
  • Artemis-413 is a valuable tool for studying DNA structural dynamics and repair mechanisms under topological stress.