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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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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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The RAG recombinase: Beyond breaking.

Chloé Lescale1, Ludovic Deriano1

  • 1Department of Immunology and Department of Genomes and Genetics, Institut Pasteur, 75015 Paris, France.

Mechanisms of Ageing and Development
|November 20, 2016
PubMed
Summary
This summary is machine-generated.

DNA double-strand breaks (DSBs) are essential for adaptive immunity via V(D)J recombination. The RAG nuclease couples breakage and repair, preventing genome disruption and potentially influencing genetic programs.

Keywords:
Adaptive immunityDNA damage responseNHEJRAG recombinaseV(D)J recombination

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Area of Science:

  • Molecular Biology
  • Immunology
  • Genetics

Background:

  • DNA double-strand breaks (DSBs) are critical DNA lesions impacting genome integrity, cancer, and aging.
  • In adaptive immunity, DSBs are essential intermediates for V(D)J recombination, generating immune receptor diversity.
  • V(D)J recombination is initiated by the lymphoid-specific RAG1 and RAG2 recombinase, forming a RAG nuclease.

Purpose of the Study:

  • To discuss how the RAG nuclease minimizes genome disruption risks during V(D)J recombination.
  • To explore the evolutionary pressure on RAG genes favoring controlled DNA repair over transposition.
  • To consider the broader impact of RAG-mediated DSBs on cellular processes and epigenetic programs.

Main Methods:

  • Review of existing literature on V(D)J recombination mechanisms.
  • Analysis of the RAG nuclease's role in coupling DNA breakage and repair.
  • Discussion of evolutionary selective pressures on RAG genes.

Main Results:

  • The RAG nuclease couples DNA breakage and repair steps in V(D)J recombination, ensuring controlled DNA end joining.
  • RAG genes, originating from a transposon, were selected to suppress transposition and promote endogenous DNA repair.
  • RAG-mediated DSBs have implications beyond V(D)J recombination, potentially affecting genetic and epigenetic landscapes.

Conclusions:

  • The RAG nuclease's function is crucial for maintaining genome stability during V(D)J recombination.
  • Evolutionary selection has shaped RAG genes to facilitate controlled DNA repair for immune diversity.
  • RAG-mediated DSBs represent a significant factor influencing cellular functions and epigenetic regulation.