Dataset Information

Assembly Pathway and Characterization of the RAG1/2-DNA Paired and Signal-end Complexes.

ABSTRACT: Mammalian immune receptor diversity is established via a unique restricted set of site-specific DNA rearrangements in lymphoid cells, known as V(D)J recombination. The lymphoid-specific RAG1-RAG2 protein complex (RAG1/2) initiates this process by binding to two types of recombination signal sequences (RSS), 12RSS and 23RSS, and cleaving at the boundaries of RSS and V, D, or J gene segments, which are to be assembled into immunoglobulins and T-cell receptors. Here we dissect the ordered assembly of the RAG1/2 heterotetramer with 12RSS and 23RSS DNAs. We find that RAG1/2 binds only a single 12RSS or 23RSS and reserves the second DNA-binding site specifically for the complementary RSS, to form a paired complex that reflects the known 12/23 rule of V(D)J recombination. The assembled RAG1/2 paired complex is active in the presence of Mg(2+), the physiologically relevant metal ion, in nicking and double-strand cleavage of both RSS DNAs to produce a signal-end complex. We report here the purification and initial crystallization of the RAG1/2 signal-end complex for atomic-resolution structure elucidation. Strict pairing of the 12RSS and 23RSS at the binding step, together with information from the crystal structure of RAG1/2, leads to a molecular explanation of the 12/23 rule.

SUBMITTER: Lapkouski M

PROVIDER: S-EPMC4505528 | biostudies-literature | 2015 Jun

REPOSITORIES: biostudies-literature

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Assembly Pathway and Characterization of the RAG1/2-DNA Paired and Signal-end Complexes.

Lapkouski Mikalai M Chuenchor Watchalee W Kim Min-Sung MS Gellert Martin M Yang Wei W

The Journal of biological chemistry 20150422 23

Mammalian immune receptor diversity is established via a unique restricted set of site-specific DNA rearrangements in lymphoid cells, known as V(D)J recombination. The lymphoid-specific RAG1-RAG2 protein complex (RAG1/2) initiates this process by binding to two types of recombination signal sequences (RSS), 12RSS and 23RSS, and cleaving at the boundaries of RSS and V, D, or J gene segments, which are to be assembled into immunoglobulins and T-cell receptors. Here we dissect the ordered assembly ...[more]

PMID: 25903130

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Dataset Information

Assembly Pathway and Characterization of the RAG1/2-DNA Paired and Signal-end Complexes.

Publications

Assembly Pathway and Characterization of the RAG1/2-DNA Paired and Signal-end Complexes.

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