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Restrictions Limiting the Generation of DNA Double Strand Breaks during Chromosomal V(D)J Recombination

Antigen receptor loci are composed of numerous variable (V), diversity (D), and joining (J) gene segments, each flanked by recombination signal sequences (RSSs). The V(D)J recombination reaction proceeds through RSS recognition and DNA cleavage steps making it possible for multiple DNA double strand...

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Autores principales: Tillman, Robert E., Wooley, Andrea L., Hughes, Maureen M., Wehrly, Tara D., Swat, Wojciech, Sleckman, Barry P.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2002
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2193590/
https://www.ncbi.nlm.nih.gov/pubmed/11828005
http://dx.doi.org/10.1084/jem.20011803
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author Tillman, Robert E.
Wooley, Andrea L.
Hughes, Maureen M.
Wehrly, Tara D.
Swat, Wojciech
Sleckman, Barry P.
author_facet Tillman, Robert E.
Wooley, Andrea L.
Hughes, Maureen M.
Wehrly, Tara D.
Swat, Wojciech
Sleckman, Barry P.
author_sort Tillman, Robert E.
collection PubMed
description Antigen receptor loci are composed of numerous variable (V), diversity (D), and joining (J) gene segments, each flanked by recombination signal sequences (RSSs). The V(D)J recombination reaction proceeds through RSS recognition and DNA cleavage steps making it possible for multiple DNA double strand breaks (DSBs) to be introduced at a single locus. Here we use ligation-mediated PCR to analyze DNA cleavage intermediates in thymocytes from mice with targeted RSS mutations at the endogenous TCRβ locus. We show that DNA cleavage does not occur at individual RSSs but rather must be coordinated between RSS pairs flanking gene segments that ultimately form coding joins. Coordination of the DNA cleavage step occurs over great distances in the chromosome and favors intra- over interchromosomal recombination. Furthermore, through several restrictions imposed on the generation of both nonpaired and paired DNA DSBs, this requirement promotes antigen receptor gene integrity and genomic stability in developing lymphocytes undergoing V(D)J recombination.
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spelling pubmed-21935902008-04-14 Restrictions Limiting the Generation of DNA Double Strand Breaks during Chromosomal V(D)J Recombination Tillman, Robert E. Wooley, Andrea L. Hughes, Maureen M. Wehrly, Tara D. Swat, Wojciech Sleckman, Barry P. J Exp Med Original Article Antigen receptor loci are composed of numerous variable (V), diversity (D), and joining (J) gene segments, each flanked by recombination signal sequences (RSSs). The V(D)J recombination reaction proceeds through RSS recognition and DNA cleavage steps making it possible for multiple DNA double strand breaks (DSBs) to be introduced at a single locus. Here we use ligation-mediated PCR to analyze DNA cleavage intermediates in thymocytes from mice with targeted RSS mutations at the endogenous TCRβ locus. We show that DNA cleavage does not occur at individual RSSs but rather must be coordinated between RSS pairs flanking gene segments that ultimately form coding joins. Coordination of the DNA cleavage step occurs over great distances in the chromosome and favors intra- over interchromosomal recombination. Furthermore, through several restrictions imposed on the generation of both nonpaired and paired DNA DSBs, this requirement promotes antigen receptor gene integrity and genomic stability in developing lymphocytes undergoing V(D)J recombination. The Rockefeller University Press 2002-02-04 /pmc/articles/PMC2193590/ /pubmed/11828005 http://dx.doi.org/10.1084/jem.20011803 Text en Copyright © 2002, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Original Article
Tillman, Robert E.
Wooley, Andrea L.
Hughes, Maureen M.
Wehrly, Tara D.
Swat, Wojciech
Sleckman, Barry P.
Restrictions Limiting the Generation of DNA Double Strand Breaks during Chromosomal V(D)J Recombination
title Restrictions Limiting the Generation of DNA Double Strand Breaks during Chromosomal V(D)J Recombination
title_full Restrictions Limiting the Generation of DNA Double Strand Breaks during Chromosomal V(D)J Recombination
title_fullStr Restrictions Limiting the Generation of DNA Double Strand Breaks during Chromosomal V(D)J Recombination
title_full_unstemmed Restrictions Limiting the Generation of DNA Double Strand Breaks during Chromosomal V(D)J Recombination
title_short Restrictions Limiting the Generation of DNA Double Strand Breaks during Chromosomal V(D)J Recombination
title_sort restrictions limiting the generation of dna double strand breaks during chromosomal v(d)j recombination
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2193590/
https://www.ncbi.nlm.nih.gov/pubmed/11828005
http://dx.doi.org/10.1084/jem.20011803
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