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 S Ciborowski, Sarah Jackson, Yue Xiong, Patrick C. Swanson

Research output: Contribution to journalArticle

21 Citations (Scopus)

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

Fingerprint

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

Keywords

  • DDB1
  • E3 ubiquitin ligase
  • RAG1
  • V(D)J recombination
  • VprBP

ASJC Scopus subject areas

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

Cite this

Kassmeier, M. D., Mondal, K., Palmer, V. L., Raval, P., Kumar, S., Perry, G. A., ... Swanson, P. C. (2012). VprBP binds full-length RAG1 and is required for B-cell development and V(D)J recombination fidelity. EMBO Journal, 31(4), 945-958. https://doi.org/10.1038/emboj.2011.455

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 S; Jackson, Sarah; Xiong, Yue; Swanson, Patrick C.

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, PS, Jackson, S, Xiong, Y & Swanson, PC 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, Michele D. ; Mondal, Koushik ; Palmer, Victoria L. ; Raval, Prafulla ; Kumar, Sushil ; Perry, Greg A. ; Anderson, Dirk K. ; Ciborowski, Pawel S ; Jackson, Sarah ; Xiong, Yue ; Swanson, Patrick C. / 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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AU - Kumar, Sushil

AU - Perry, Greg A.

AU - Anderson, Dirk K.

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AU - Swanson, Patrick C.

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