Full-length RAG-2, and Not Full-length RAG-1, Specifically Suppresses RAG-mediated Transposition but Not Hybrid Joint Formation or Disintegration

Patrick Swanson, Dustin Volkmer, Lei Wang

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

RAG-1 and RAG-2 initiate V(D)J recombination by introducing DNA breaks at recombination signal sequences flanking a pair of antigen receptor gene segments. Occasionally, the RAG proteins mediate two other alternative DNA rearrangements in vivo: the rejoining of signal and coding ends and the transposition of signal ends into unrelated DNA. In contrast, truncated, catalytically active "core" RAG proteins readily catalyze these reactions in vitro, suggesting that full-length RAG proteins directly or indirectly suppress these undesired reactions in vivo. To discriminate between direct and indirect suppression models, full-length RAG proteins were purified and characterized in vitro. From mammalian cells, full-length RAG-1 is readily purified with core RAG-2 but not full-length RAG-2 and vice versa. Despite differences in DNA binding activity, recombinase containing either core or full-length RAG-1 or RAG-2 possess comparable cleavage, rejoining, and end-processing activity, as well as similar usage preferences for canonical versus cryptic recombination signals. However, recombinase containing full-length RAG-2, but not full-length RAG-1, exhibits dramatically reduced transposition activity in vitro. These data suggest RAG-mediated transposition and rejoining are differentially regulated by the full-length RAG proteins in vivo (the former directly by RAG-2 and the latter indirectly through other factors) and argue that non-core portions of the RAG proteins have little or no direct influence over V(D)J recombinase site specificity.

Original languageEnglish
Pages (from-to)4034-4044
Number of pages11
JournalJournal of Biological Chemistry
Volume279
Issue number6
DOIs
StatePublished - Feb 6 2004

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Disintegration
Joints
Recombinases
Proteins
DNA
Genetic Recombination
VDJ Recombinases
V(D)J Recombination
Antigen Receptors
DNA Breaks
Gene Rearrangement
Protein Sorting Signals
Genes
Cells
Processing
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Full-length RAG-2, and Not Full-length RAG-1, Specifically Suppresses RAG-mediated Transposition but Not Hybrid Joint Formation or Disintegration. / Swanson, Patrick; Volkmer, Dustin; Wang, Lei.

In: Journal of Biological Chemistry, Vol. 279, No. 6, 06.02.2004, p. 4034-4044.

Research output: Contribution to journalArticle

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