VprBP (DCAF1) regulates RAG1 expression independently of dicer by mediating RAG1 degradation

N. Max Schabla, Greg A. Perry, Victoria L. Palmer, Patrick Swanson

Research output: Contribution to journalArticle

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Abstract

The assembly of Ig genes in developing B lymphocytes by V(D)J recombination is initiated by the RAG1-RAG2 endonuclease complex. We previously identified an interaction between RAG1 and viral protein R binding protein (VprBP) (also known as DNA damage binding protein 1 cullin 4-associated factor 1 [DCAF1]), a substrate receptor for the cullin 4-really interesting new gene (RING) E3 ubiquitin ligase (CRL4). We report in this article that in mice, B cell-intrinsic loss of VprBP increases RAG1 protein levels and disrupts expression of the endoribonuclease Dicer, which is essential for microRNA maturation. Rag1/2 transcription is known to be derepressed by loss of microRNA-mediated suppression of phosphatase and tensin homolog, raising the possibility that the elevated level of RAG1 observed in VprBP-deficient B cells is caused indirectly by the loss of Dicer. However, we show that VprBP restrains RAG1 expression posttranscriptionally and independently of Dicer. Specifically, loss of VprBP stabilizes RAG1 protein, which we show is normally degraded via a mechanism requiring both 20S proteasome and cullin-RING E3 ubiquitin ligase activity. Furthermore, we show that RAG1 stabilization through small molecule inhibition of cullin-RING E3 ubiquitin ligase activation promotes V(D)J recombination in a murine pre-B cell line. Thus, in addition to identifying a role for VprBP in maintaining Dicer levels in B cells, our findings reveal the basis for RAG1 turnover and provide evidence that the CRL4VprBP(DCAF1) complex functions to maintain physiological levels of V(D)J recombination.

Original languageEnglish (US)
Pages (from-to)930-939
Number of pages10
JournalJournal of Immunology
Volume201
Issue number3
DOIs
StatePublished - Aug 1 2018

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vpr Gene Products
Protein Binding
Carrier Proteins
Cullin Proteins
V(D)J Recombination
Ubiquitin-Protein Ligases
B-Lymphocytes
MicroRNAs
Endoribonucleases
Genes
Immunoglobulin Genes
B-Lymphoid Precursor Cells
Endonucleases
DNA-Binding Proteins
Proteasome Endopeptidase Complex
Cullin 1
Phosphoric Monoester Hydrolases
DNA Damage
Proteins
Cell Line

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

VprBP (DCAF1) regulates RAG1 expression independently of dicer by mediating RAG1 degradation. / Max Schabla, N.; Perry, Greg A.; Palmer, Victoria L.; Swanson, Patrick.

In: Journal of Immunology, Vol. 201, No. 3, 01.08.2018, p. 930-939.

Research output: Contribution to journalArticle

Max Schabla, N. ; Perry, Greg A. ; Palmer, Victoria L. ; Swanson, Patrick. / VprBP (DCAF1) regulates RAG1 expression independently of dicer by mediating RAG1 degradation. In: Journal of Immunology. 2018 ; Vol. 201, No. 3. pp. 930-939.
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