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  1. Artikel: Mechanisms that direct ordered assembly of T cell receptor beta locus V, D, and J gene segments.

    Sleckman, B P / Bassing, C H / Hughes, M M / Okada, A / D'Auteuil, M / Wehrly, T D / Woodman, B B / Davidson, L / Chen, J / Alt, F W

    Proceedings of the National Academy of Sciences of the United States of America

    2000  Band 97, Heft 14, Seite(n) 7975–7980

    Abstract: T cell receptor (TCR) beta variable region genes are assembled in progenitor T cells from germ-line Vbeta, Dbeta, and Jbeta segments via an ordered two-step process in which Dbeta to Jbeta rearrangements occur on both alleles before appendage of a Vbeta ... ...

    Abstract T cell receptor (TCR) beta variable region genes are assembled in progenitor T cells from germ-line Vbeta, Dbeta, and Jbeta segments via an ordered two-step process in which Dbeta to Jbeta rearrangements occur on both alleles before appendage of a Vbeta to a preexisting DJbeta complex. Direct joining of Vbeta segments to nonrearranged Dbeta or Jbeta segments, while compatible with known restrictions on the V(D)J recombination mechanism, are infrequent within the endogenous TCRbeta locus. We have analyzed mechanisms that mediate ordered Vbeta, Dbeta, and Jbeta assembly via an approach in which TCRbeta minilocus recombination substrates were introduced into embryonic stem cells and then analyzed for rearrangement in normal thymocytes by recombinase-activating gene 2-deficient blastocyst complementation. These analyses demonstrated that Vbeta segments are preferentially targeted for rearrangement to Dbeta as opposed to Jbeta segments. In addition, we further demonstrated that Vbeta segments can be appended to nonrearranged endogenous Dbeta segments in which we have eliminated the ability of Dbeta segments to join to Jbeta segments. Our findings are discussed in the context of the mechanisms that regulate the ordered assembly and utilization of V, D, and J segments.
    Mesh-Begriff(e) Animals ; Cell Lineage ; Chimera ; DNA-Binding Proteins/genetics ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; Genes, T-Cell Receptor beta ; Genetic Complementation Test ; Mice ; Models, Genetic ; Receptors, Antigen, T-Cell, alpha-beta/genetics ; Recombination, Genetic
    Chemische Substanzen DNA-Binding Proteins ; Rag2 protein, mouse ; Receptors, Antigen, T-Cell, alpha-beta ; V(D)J recombination activating protein 2
    Sprache Englisch
    Erscheinungsdatum 2000-07-05
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.130190597
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  2. Artikel: Recombination signal sequences restrict chromosomal V(D)J recombination beyond the 12/23 rule.

    Bassing, C H / Alt, F W / Hughes, M M / D'Auteuil, M / Wehrly, T D / Woodman, B B / Gärtner, F / White, J M / Davidson, L / Sleckman, B P

    Nature

    2000  Band 405, Heft 6786, Seite(n) 583–586

    Abstract: The genes encoding the variable regions of lymphocyte antigen receptors are assembled from variable (V), diversity (D) and joining (J) gene segments. V(D)J recombination is initiated by the recombinase activating gene (RAG)-1 and -2 proteins, which ... ...

    Abstract The genes encoding the variable regions of lymphocyte antigen receptors are assembled from variable (V), diversity (D) and joining (J) gene segments. V(D)J recombination is initiated by the recombinase activating gene (RAG)-1 and -2 proteins, which introduce DNA double-strand breaks between the V, D and J segments and their flanking recombination signal sequences (RSSs). Generally expressed DNA repair proteins then carry out the joining reaction. The conserved heptamer and nonamer sequences of the RSSs are separated by non-conserved spacers of 12 or 23 base pairs (forming 12-RSSs and 23-RSSs). The 12/23 rule, which is mediated at the level of RAG-1/2 recognition and cutting, specifies that V(D)J recombination occurs only between a gene segment flanked by a 12-RSS and one flanked by a 23-RSS. Vbeta segments are appended to DJbeta rearrangements, with little or no direct Vbeta to Jbeta joining, despite 12/23 compatibility of Vbeta 23-RSSs and Jbeta12-RSSs. Here we use embryonic stem cells and mice with a modified T-cell receptor (TCR)beta locus containing only one Dbeta (Dbeta1) gene segment and one Jbeta (Jbeta1) gene cluster to show that the 5' Dbeta1 12-RSS, but not the Jbeta1 12-RSSs, targets rearrangement of a diverse Vbeta repertoire. This targeting is precise and position-independent. This additional restriction on V(D)J recombination has important implications for the regulation of variable region gene assembly and repertoire development.
    Mesh-Begriff(e) Alleles ; Animals ; Cell Line ; Chimera ; Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor ; Hybridomas ; Mice ; Multigene Family ; Mutagenesis ; Receptors, Antigen, T-Cell, alpha-beta/genetics ; Recombination, Genetic ; Stem Cells
    Chemische Substanzen Receptors, Antigen, T-Cell, alpha-beta
    Sprache Englisch
    Erscheinungsdatum 2000-06-01
    Erscheinungsland England
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 120714-3
    ISSN 1476-4687 ; 0028-0836
    ISSN (online) 1476-4687
    ISSN 0028-0836
    DOI 10.1038/35014635
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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