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PMID: 23964096 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

E3-ubiquitin ligase Nedd4 determines the fate of AID-associated RNA polymerase II in B cells.

Genes & development ·Vol. 27 ·No. 16 ·2013-08-15 ·Pages 1821-33

Sun J, Keim CD, Wang J, Kazadi D, Oliver PM, Rabadan R, Basu U

Abstract

Programmed mutagenesis of the immunoglobulin locus of B lymphocytes during class switch recombination (CSR) and somatic hypermutation requires RNA polymerase II (polII) transcription complex-dependent targeting of the DNA mutator activation-induced cytidine deaminase (AID). AID deaminates cytidine residues on substrate sequences in the immunoglobulin (Ig) locus via a transcription-dependent mechanism, and this activity is stimulated by the RNA polII stalling cofactor Spt5 and the 11-subunit cellular noncoding RNA 3'-5' exonucleolytic processing complex RNA exosome. The mechanism by which the RNA exosome recognizes immunoglobulin locus RNA substrates to stimulate AID DNA deamination activity on its in vivo substrate sequences is an important question. Here we report that E3-ubiquitin ligase Nedd4 destabilizes AID-associated RNA polII by a ubiquitination event, leading to generation of 3' end free RNA exosome RNA substrates at the Ig locus and other AID target sequences genome-wide. We found that lack of Nedd4 activity in B cells leads to accumulation of RNA exosome substrates at AID target genes and defective CSR. Taken together, our study links noncoding RNA processing following RNA polII pausing with regulation of the mutator AID protein. Our study also identifies Nedd4 as a regulator of noncoding RNAs that are generated by stalled RNA polII genome-wide.

Keywords
Nedd4 RNA polymerase II stalling RNA polymerase II ubiquitination activation-induced deaminase immunoglobulin locus transcription noncoding RNA
MeSH Terms
Animals B-Lymphocytes/enzymology Cell Line, Tumor Cells, Cultured Cytidine Deaminase/metabolism Endosomal Sorting Complexes Required for Transport/genetics,metabolism Exosome Multienzyme Ribonuclease Complex/metabolism Gene Knockdown Techniques HEK293 Cells Humans Immunoglobulin Switch Region/genetics Mice Nedd4 Ubiquitin Protein Ligases Nuclear Proteins/metabolism Protein Binding RNA Polymerase II/metabolism Transcriptional Elongation Factors/metabolism Ubiquitin-Protein Ligases/genetics,metabolism Ubiquitination
Chemicals
Endosomal Sorting Complexes Required for Transport Nuclear Proteins SUPT5H protein, human Transcriptional Elongation Factors Nedd4 Ubiquitin Protein Ligases Nedd4 protein, human Nedd4l protein, mouse Ubiquitin-Protein Ligases RNA Polymerase II Exosome Multienzyme Ribonuclease Complex AICDA (activation-induced cytidine deaminase) Cytidine Deaminase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Sun Jianbo
Department of Microbiology and Immunology, Columbia University, New York, New York 10032, USA.
Keim Celia D
Wang Jiguang
Kazadi David
Oliver Paula M
Rabadan Raul
Basu Uttiya
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
1549-5477
DOI
10.1101/gad.210211.112
Published
2013-08-15
Pages
1821-33
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC3759698
Subset
IM
Grants
NIAID NIH HHS · R01 AI099195 · United States
NIH HHS · 1DP2OD008651-01 · United States
NIAID NIH HHS · R01 AI093566 · United States
NIAID NIH HHS · 1R01AI099195-01A1 · United States
NIAID NIH HHS · F31AI098411-01A1 · United States
NIAID NIH HHS · F31 AI098411 · United States
NIH HHS · DP2 OD008651 · United States
External Links
PubMed source DOI: 10.1101/gad.210211.112 PMC Full Text View journal details
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