Leaky severe combined immunodeficiency and aberrant DNA rearrangements due to a hypomorphic RAG1 mutation

Blood. 2009 Mar 26;113(13):2965-75. doi: 10.1182/blood-2008-07-165167. Epub 2009 Jan 6.

Abstract

The RAG1/2 endonuclease initiates programmed DNA rearrangements in progenitor lymphocytes by generating double-strand breaks at specific recombination signal sequences. This process, known as V(D)J recombination, assembles the vastly diverse antigen receptor genes from numerous V, D, and J coding segments. In vitro biochemical and cellular transfection studies suggest that RAG1/2 may also play postcleavage roles by forming complexes with the recombining ends to facilitate DNA end processing and ligation. In the current study, we examine the in vivo consequences of a mutant form of RAG1, RAG1-S723C, that is proficient for DNA cleavage, yet exhibits defects in postcleavage complex formation and end joining in vitro. We generated a knockin mouse model harboring the RAG1-S723C hypomorphic mutation and examined the immune system in this fully in vivo setting. RAG1-S723C homozygous mice exhibit impaired lymphocyte development and decreased V(D)J rearrangements. Distinct from RAG nullizygosity, the RAG1-S723C hypomorph results in aberrant DNA double-strand breaks within rearranging loci. RAG1-S723C also predisposes to thymic lymphomas associated with chromosomal translocations in a p53 mutant background, and heterozygosity for the mutant allele accelerates age-associated immune system dysfunction. Thus, our study provides in vivo evidence that implicates aberrant RAG1/2 activity in lymphoid tumor development and premature immunosenescence.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aging / genetics
  • Aging / immunology
  • Amino Acid Substitution / physiology
  • Animals
  • B-Lymphocytes / immunology
  • B-Lymphocytes / pathology
  • Gene Knock-In Techniques
  • Gene Rearrangement / genetics*
  • Homeodomain Proteins / genetics*
  • Homozygote
  • Lymphoma / genetics
  • Lymphoma / immunology
  • Mice
  • Mice, Transgenic
  • Mutation, Missense* / physiology
  • Phenotype
  • Severe Combined Immunodeficiency / genetics*
  • Severe Combined Immunodeficiency / immunology
  • Severe Combined Immunodeficiency / pathology
  • T-Lymphocytes / immunology
  • T-Lymphocytes / pathology
  • Thymus Neoplasms / genetics
  • Thymus Neoplasms / immunology
  • VDJ Exons

Substances

  • Homeodomain Proteins
  • RAG-1 protein