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Ras orchestrates exit from the cell cycle and light-chain recombination during early B cell development

Nature Immunology volume 10, pages 1110–1117 (2009)Cite this article

Abstract

Signals through the pre–B cell antigen receptor (pre-BCR) and interleukin 7 receptor (IL-7R) coordinate pre–B cell population expansion with subsequent recombination of the locus encoding immunoglobulin κ-chain (Igk). Although many 'downstream' effectors of each receptor are known, how they integrate to mediate development has remained unclear. Here we report that pre-BCR-mediated activation of the Ras-MEK-Erk signaling pathway silenced transcription of Ccnd3 (encoding cyclin D3) and coordinated exit from the cell cycle with induction of the transcription factor E2A and the initiation of Igk recombination. IL-7R-mediated activation of the transcription factor STAT5 opposed this pathway by promoting Ccnd3 expression and concomitantly inhibiting Igk transcription by binding to the Igk intronic enhancer and preventing E2A recruitment. Our data show how pre-BCR signaling poises pre–B cells to undergo differentiation after escape from IL-7R signaling.

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Figure 1: MEK-Erk regulate both cyclins and the Igk recombination machinery.
Figure 2: Ras-MEK activation required for exit from the cell cycle after IL-7 withdrawal.
Figure 3: Ras-MEK activation is required for the induction of Igk recombination after IL-7 withdrawal.
Figure 4: Aiolos is induced in pre–B cells and can suppress Ccnd3 transcription.
Figure 5: Downstream of Ras, Aiolos is required for suppression of Ccnd3 and exit from the cell cycle after IL-7 withdrawal.
Figure 6: Downregulation of Id3 and upregulation of E2A by Ras-MEK signaling increases E2A binding activity.
Figure 7: More Igk transcription and rearrangement in Id3−/− mice.
Figure 8: STAT5 regulates Igk transcription and binds directly to Eκi.

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Acknowledgements

We thank T. Gajewski (University of Chicago) for cDNA encoding CA-Ras, CA-STAT5B and DN-Ras; N.G. Ahn (University of Colorado, Boulder) for cDNA encoding DN-MEK; M. Veselits for technical assistance; R. Duggan and D. Leclerc for cell-sorting services; and F. Gounari for discussions.

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Authors and Affiliations

  1. Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, Illinois, USA

    Malay Mandal, Sarah E Powers & Marcus R Clark

  2. Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, Illinois, USA

    Kyoko Ochiai & Harinder Singh

  3. Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts, USA

    Katia Georgopoulos

  4. Department of Pathology, University of Chicago, Chicago, Illinois, USA

    Barbara L Kee

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Contributions

M.M. designed, did, interpreted and prepared figures of most experiments and assisted in preparing the initial manuscript; S.E.P. assisted in the design and analysis of many experiments; K.O. assisted with chromatin immunoprecipitation; K.G. provided Aiolos-specific reagents and scientific advice and assisted in preparing the manuscript; B.L.K. provided E2A-specific reagents, contributed to the design of some experiments, provided scientific advice and edited the manuscript; H.S. assisted in establishing many of the experimental systems, provided substantial scientific input and edited the final manuscript; and M.R.C. oversaw the entire project and prepared the final manuscript.

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Correspondence to Marcus R Clark.

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Mandal, M., Powers, S., Ochiai, K. et al. Ras orchestrates exit from the cell cycle and light-chain recombination during early B cell development. Nat Immunol 10, 1110–1117 (2009). https://doi.org/10.1038/ni.1785

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