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How mouse RAG recombinase avoids DNA transposition.

Xuemin Chen1, Yanxiang Cui2, Huaibin Wang1

  • 1Laboratory of Molecular Biology, NIDDK, National Institutes of Health, Bethesda, MD, USA.

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The RAG1-RAG2 recombinase (RAG) enzyme

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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Immunology

Background:

  • The RAG1-RAG2 recombinase (RAG) is crucial for V(D)J recombination in lymphocytes.
  • RAG shares homology with the RNH-type transposase family, suggesting potential for DNA transposition.
  • Understanding RAG's regulation is key to preventing aberrant DNA cleavage and transposition.

Purpose of the Study:

  • To elucidate the structural mechanism by which RAG-catalyzed transposition is inhibited in developing lymphocytes.
  • To investigate the role of RAG2 in modulating target DNA structure during transpositional complexes.

Main Methods:

  • Determined the 3.1-Å resolution crystal structure of a mouse RAG DNA-strand transfer complex.
  • Analyzed the structural features of the target DNA (T-form) and its interaction with RAG.

Main Results:

  • The target DNA adopts a T-form with two sharp kinks (>80°) near the integration site.
  • RAG2 enforces these kinks and inter-segment twisting, attenuating unwanted transposition.
  • Unlike other transposases, the key kink in RAG complexes is 1 bp away from the integration site, facilitating DNA release.

Conclusions:

  • RAG2 plays a critical role in regulating RAG transposition by inducing specific DNA distortions.
  • The structural mechanism elucidated explains how RAG efficiently catalyzes DNA cleavage and promotes V(D)J recombination while suppressing unwanted transposition.
  • This regulation is essential for lymphocyte development and genomic integrity.