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The splicing regulator PTBP2 interacts with the cytidine deaminase AID and promotes binding of AID to switch-region DNA

Nature Immunology volume 12, pages 160–166 (2011)Cite this article

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Abstract

During immunoglobulin class-switch recombination (CSR), the cytidine deaminase AID induces double-strand breaks into transcribed, repetitive DNA elements called switch sequences. The mechanism that promotes the binding of AID specifically to switch regions remains to be elucidated. Here we used a proteomic screen with in vivo biotinylation of AID to identify the splicing regulator PTBP2 as a protein that interacts with AID. Knockdown of PTBP2 mediated by short hairpin RNA in B cells led to a decrease in binding of AID to transcribed switch regions, which resulted in considerable impairment of CSR. PTBP2 is thus an effector of CSR that promotes the binding of AID to switch-region DNA.

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Figure 1: AID interacts with PTBP2.
Figure 2: Knockdown of PTBP2 impairs CSR.
Figure 3: Germline transcription is unaffected in cells with knockdown of PTBP2.
Figure 4: The expression and nuclear localization of AID are not altered in cells with knockdown of PTBP2.
Figure 5: PTBP2 binds S transcripts in vitro.
Figure 6: PTBP2 promotes the binding of AID to S-region DNA.
Figure 7: Knockdown of PTBP2 impairs CSR and the binding of AID to activated S regions in primary B cells.

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Acknowledgements

We thank T. Honjo (University of Kyoto) for AID-deficient (Aicda−/−) mice and CH12 cells; S. Orkin (Harvard Medical School) for the biotag vector and BirA plasmids and all members of the Chaudhuri laboratory for discussions and suggestions. Supported by the Damon Runyon Cancer Research Fund (J.C.), the Alfred Bressler Foundation (J.C.), the National Institutes of Health (J.C.) and the Cancer Research Institute (U.N.).

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Authors and Affiliations

  1. Graduate Program in Immunology and Microbial Pathogenesis, Weill-Cornell Medical School, New York, New York, USA

    Urszula Nowak, Allysia J Matthews, Simin Zheng & Jayanta Chaudhuri

  2. Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, USA

    Jayanta Chaudhuri

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  1. Urszula Nowak
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  2. Allysia J Matthews
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Contributions

U.N. and J.C. designed and did experiments, analyzed the data and wrote the manuscript; and A.J.M. did the shRNA knockdown for the AID-localization studies and with S.Z. established shRNA knockdown techniques in CH12 cells.

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Correspondence to Jayanta Chaudhuri.

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The authors declare no competing financial interests.

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Nowak, U., Matthews, A., Zheng, S. et al. The splicing regulator PTBP2 interacts with the cytidine deaminase AID and promotes binding of AID to switch-region DNA. Nat Immunol 12, 160–166 (2011). https://doi.org/10.1038/ni.1977

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