A stable RAG1-RAG2-DNA complex that is active in V(D)J cleavage

Kevin Hiom, Martin Gellert

    Research output: Contribution to journalArticle

    179 Citations (Scopus)

    Abstract

    The RAG1 and RAG2 proteins initiate V(D)J recombination by making specific double-strand DNA breaks at recombination signal sequences. We show here that RAG1 and RAG2 bind specifically to this sequence, forming a stable protein-DNA complex. The complex requires the conserved heptamer and nonamer motifs of the recombination signal as well as both the RAG1 and RAG2 proteins. This complex is able to either nick or form hairpins at the V(D)J signal sequence, depending on the divalent cation present. A complex trapped using Ca2+ is subsequently active when transferred to Mg2+ or Mn2+. After cleavage, the complex is destabilized and the RAG proteins dissociate. We term this early precursor in the V(D)J recombination reaction a "stable cleavage complex."

    Original languageEnglish
    Pages (from-to)65-72
    Number of pages8
    JournalCell
    Volume88
    Issue number1
    DOIs
    Publication statusPublished - 10 Jan 1997

    Fingerprint

    V(D)J Recombination
    DNA
    Protein Sorting Signals
    Genetic Recombination
    Proteins
    Double-Stranded DNA Breaks
    Divalent Cations

    Keywords

    • Cations, Divalent
    • DNA
    • DNA-binding proteins
    • Homeodomain proteins
    • Oligodeoxyribonucleotides
    • Protein binding
    • Proteins
    • Recombination, Genetic

    Cite this

    Hiom, Kevin ; Gellert, Martin. / A stable RAG1-RAG2-DNA complex that is active in V(D)J cleavage. In: Cell. 1997 ; Vol. 88, No. 1. pp. 65-72.
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    A stable RAG1-RAG2-DNA complex that is active in V(D)J cleavage. / Hiom, Kevin; Gellert, Martin.

    In: Cell, Vol. 88, No. 1, 10.01.1997, p. 65-72.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - A stable RAG1-RAG2-DNA complex that is active in V(D)J cleavage

    AU - Hiom, Kevin

    AU - Gellert, Martin

    PY - 1997/1/10

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    N2 - The RAG1 and RAG2 proteins initiate V(D)J recombination by making specific double-strand DNA breaks at recombination signal sequences. We show here that RAG1 and RAG2 bind specifically to this sequence, forming a stable protein-DNA complex. The complex requires the conserved heptamer and nonamer motifs of the recombination signal as well as both the RAG1 and RAG2 proteins. This complex is able to either nick or form hairpins at the V(D)J signal sequence, depending on the divalent cation present. A complex trapped using Ca2+ is subsequently active when transferred to Mg2+ or Mn2+. After cleavage, the complex is destabilized and the RAG proteins dissociate. We term this early precursor in the V(D)J recombination reaction a "stable cleavage complex."

    AB - The RAG1 and RAG2 proteins initiate V(D)J recombination by making specific double-strand DNA breaks at recombination signal sequences. We show here that RAG1 and RAG2 bind specifically to this sequence, forming a stable protein-DNA complex. The complex requires the conserved heptamer and nonamer motifs of the recombination signal as well as both the RAG1 and RAG2 proteins. This complex is able to either nick or form hairpins at the V(D)J signal sequence, depending on the divalent cation present. A complex trapped using Ca2+ is subsequently active when transferred to Mg2+ or Mn2+. After cleavage, the complex is destabilized and the RAG proteins dissociate. We term this early precursor in the V(D)J recombination reaction a "stable cleavage complex."

    KW - Cations, Divalent

    KW - DNA

    KW - DNA-binding proteins

    KW - Homeodomain proteins

    KW - Oligodeoxyribonucleotides

    KW - Protein binding

    KW - Proteins

    KW - Recombination, Genetic

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