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The influence of transcriptional orientation on endogenous switch region function

Nature Immunology volume 4, pages 435–441 (2003)Cite this article

Abstract

Immunoglobulin heavy chain (IgH) class switch recombination (CSR) takes place between large switch (S) regions that precede exons of the constant region. The precise functions of the S region are controversial, although transcription of the S region targets CSR. We have tested the effects of deletion, inversion and replacement of the endogenous 12-kilobase Sγ1 region on CSR in vivo. Here we show that Sγ1 is required for CSR, that CSR is effected by a 1-kilobase sequence that generates a G-rich transcript, and that inversion of Sγ1 or the G-rich sequence decreases CSR. We conclude that Sγ1 function is dependent on orientation and lacks an absolute requirement for common S region motifs. We propose that single-stranded DNA stabilized by transcription-dependent, higher order structures is a primary substrate of CSR.

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Figure 1: Mutation of Sγ1.
Figure 2: RT-PCR analysis of γ1 germline transcripts.
Figure 3: ELISA analysis of IgG1 production in Sγ1 mutants.
Figure 4: Location of μ-γ1 recombination junctions.

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Acknowledgements

We thank L. Davidson, D. Foy, A. Williams, N. Stokes and C. Kennedy for mouse work, and J. Manis, C. Bassing, A. Zarrin and J. Chaudhuri for comments on the manuscript. This work was supported by US National Institutes of Health grant AI31541 (to F.W.A.) and NIH training grant AI07512 (to M.T.). K.C. is supported by a Pfizer Postdoctoral Fellowship in Rheumatology/Immunology. F.W.A is an Investigator and R.S. is an Associate of the Howard Hughes Medical Institute.

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  1. Reiko Shinkura and Ming Tian: These authors contributed equally to this work.

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  1. Howard Hughes Medical Institute, The Children's Hospital, The Center for Blood Research, and Department of Genetics, Harvard University Medical School, Boston, 02115, MA, USA

    Reiko Shinkura, Ming Tian, Michele Smith, Katrin Chua, Yuko Fujiwara & Frederick W. Alt

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Shinkura, R., Tian, M., Smith, M. et al. The influence of transcriptional orientation on endogenous switch region function. Nat Immunol 4, 435–441 (2003). https://doi.org/10.1038/ni918

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