Abstract
V(D)J recombination, the rearrangement of gene segments to assemble Ig and T cell receptor coding regions, is vital to B and T lymphocyte development. Here, we demonstrate that the V(D)J recombinase protein RAG1 undergoes ubiquitylation in cells. In vitro, the RING finger domain of RAG1 acts as a ubiquitin ligase that mediates its own ubiquitylation at a highly conserved K residue in the RAG1 amino-terminal region. Ubiquitylation is best supported by a specific ubiquitin-conjugating enzyme, UbcH3/CDC34, and requires an intact RAG1 RING finger motif. Disruption of the RING finger and certain RAG1 N-terminal truncations are associated with immunodeficiency in human patients, suggesting that RAG1's ubiquitin ligase is required for its biological role in lymphocyte development.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Base Sequence
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Binding Sites
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Cloning, Molecular
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DNA Primers
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / metabolism
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Homeodomain Proteins / chemistry
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Homeodomain Proteins / genetics
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Homeodomain Proteins / metabolism*
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Humans
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Kinetics
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Mice
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Nuclear Proteins
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Polymerase Chain Reaction
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Sequence Alignment
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Sequence Homology, Amino Acid
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Ubiquitin / metabolism*
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VDJ Recombinases / metabolism*
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Zinc / metabolism
Substances
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DNA Primers
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DNA-Binding Proteins
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Homeodomain Proteins
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Nuclear Proteins
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RAG2 protein, human
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Rag2 protein, mouse
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Recombinant Proteins
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Ubiquitin
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V(D)J recombination activating protein 2
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RAG-1 protein
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VDJ Recombinases
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Zinc