VprBP binds full-length RAG1 and is required for B-cell development and V(D)J recombination fidelity

Michele D. Kassmeier, Koushik Mondal, Victoria L. Palmer, Prafulla Raval, Sushil Kumar, Greg A. Perry, Dirk K. Anderson, Pawel Ciborowski, Sarah Jackson, Yue Xiong, Patrick Swanson

Research output: Contribution to journalArticle

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Abstract

The N-terminus of full-length RAG1, though dispensable for RAG1/2 cleavage activity, is required for efficient V(D)J recombination. This region supports RING E3 ubiquitin ligase activity in vitro, but whether full-length RAG1 functions as a single subunit or a multi-subunit E3 ligase in vivo is unclear. We show the multi-subunit cullin RING E3 ligase complex VprBP/DDB1/Cul4A/Roc1 associates with full-length RAG1 through VprBP. This complex is assembled into RAG protein-DNA complexes, and supports in-vitro ubiquitylation activity that is insensitive to RAG1 RING domain mutations. Conditional B lineage-specific VprBP disruption arrests B-cell development at the pro-B-to-pre-B cell transition, but this block is bypassed by expressing rearranged immunoglobulin transgenes. Mice with a conditional VprBP disruption show modest reduction of D-J H rearrangement, whereas V H-DJ H and V κ-J κ rearrangements are severely impaired. D-J H coding joints from VprBP-insufficent mice show longer junctional nucleotide insertions and a higher mutation frequency in D and J segments than normal. These data suggest full-length RAG1 recruits a cullin RING E3 ligase complex to ubiquitylate an unknown protein(s) to limit error-prone repair during V(D)J recombination.

Original languageEnglish
Pages (from-to)945-958
Number of pages14
JournalEMBO Journal
Volume31
Issue number4
DOIs
StatePublished - Feb 15 2012

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V(D)J Recombination
Ubiquitin-Protein Ligases
B-Lymphocytes
Cullin Proteins
Cells
B-Lymphoid Precursor Cells
Ubiquitination
Mutation Rate
Transgenes
Immunoglobulins
Proteins
Repair
Nucleotides
Joints
Mutation
DNA
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

VprBP binds full-length RAG1 and is required for B-cell development and V(D)J recombination fidelity. / Kassmeier, Michele D.; Mondal, Koushik; Palmer, Victoria L.; Raval, Prafulla; Kumar, Sushil; Perry, Greg A.; Anderson, Dirk K.; Ciborowski, Pawel; Jackson, Sarah; Xiong, Yue; Swanson, Patrick.

In: EMBO Journal, Vol. 31, No. 4, 15.02.2012, p. 945-958.

Research output: Contribution to journalArticle

Kassmeier, MD, Mondal, K, Palmer, VL, Raval, P, Kumar, S, Perry, GA, Anderson, DK, Ciborowski, P, Jackson, S, Xiong, Y & Swanson, P 2012, 'VprBP binds full-length RAG1 and is required for B-cell development and V(D)J recombination fidelity', EMBO Journal, vol. 31, no. 4, pp. 945-958. https://doi.org/10.1038/emboj.2011.455
Kassmeier MD, Mondal K, Palmer VL, Raval P, Kumar S, Perry GA et al. VprBP binds full-length RAG1 and is required for B-cell development and V(D)J recombination fidelity. EMBO Journal. 2012 Feb 15;31(4):945-958. https://doi.org/10.1038/emboj.2011.455
Kassmeier, Michele D. ; Mondal, Koushik ; Palmer, Victoria L. ; Raval, Prafulla ; Kumar, Sushil ; Perry, Greg A. ; Anderson, Dirk K. ; Ciborowski, Pawel ; Jackson, Sarah ; Xiong, Yue ; Swanson, Patrick. / VprBP binds full-length RAG1 and is required for B-cell development and V(D)J recombination fidelity. In: EMBO Journal. 2012 ; Vol. 31, No. 4. pp. 945-958.
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