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 language | English |
|---|---|
| Pages (from-to) | 945-958 |
| Number of pages | 14 |
| Journal | EMBO Journal |
| Volume | 31 |
| Issue number | 4 |
| DOIs | |
| State | Published - Feb 15 2012 |
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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 journal › Article
}
TY - JOUR
T1 - VprBP binds full-length RAG1 and is required for B-cell development and V(D)J recombination fidelity
AU - Kassmeier, Michele D.
AU - Mondal, Koushik
AU - Palmer, Victoria L.
AU - Raval, Prafulla
AU - Kumar, Sushil
AU - Perry, Greg A.
AU - Anderson, Dirk K.
AU - Ciborowski, Pawel
AU - Jackson, Sarah
AU - Xiong, Yue
AU - Swanson, Patrick
PY - 2012/2/15
Y1 - 2012/2/15
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84857059697&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84857059697&partnerID=8YFLogxK
U2 - 10.1038/emboj.2011.455
DO - 10.1038/emboj.2011.455
M3 - Article
C2 - 22157821
AN - SCOPUS:84857059697
VL - 31
SP - 945
EP - 958
JO - EMBO Journal
JF - EMBO Journal
SN - 0261-4189
IS - 4
ER -