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SLP-65 regulates immunoglobulin light chain gene recombination through the PI(3)K-PKB-Foxo pathway

Nature Immunology volume 9, pages 623–631 (2008)Cite this article

Abstract

Although the essential role of the adaptor protein SLP-65 in pre-B cell differentiation is established, the molecular mechanism underlying its function is poorly understood. In this study, we uncover a link between SLP-65–dependent signaling and the phosphoinositide-3-OH kinase (PI(3)K)–protein kinase B (PKB)–Foxo pathway. We show that the forkhead box transcription factor Foxo3a promotes light chain rearrangement in pre-B cells. Our data suggest that PKB suppresses light chain recombination by phosphorylating Foxo proteins, whereas reconstitution of SLP-65 function counteracts PKB activation and promotes Foxo3a and Foxo1 activity in pre-B cells. Together, these data illuminate a molecular function of SLP-65 and identify a key role for Foxo proteins in the regulation of light chain recombination, receptor editing and B cell selection.

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Figure 1: Signaling through PI(3)K and PKB regulates light chain recombination in pre-B cells.
Figure 2: PKB targets Foxo3a in pre-B cells.
Figure 3: Foxo3a-A3 promotes differentiation of pre-B cells.
Figure 4: Foxo3a-A3 prolongs the G1 phase of the cell cycle and stabilizes RAG-2.
Figure 5: Foxo3a-A3 counteracts the block in κ light chain expression in pre-B cells mediated by constitutively active PKB.
Figure 6: Foxo3a-deficient pre-B cells show impaired differentiation into BCR+ cells in vitro.
Figure 7: SLP-65 activates Foxo3a and Foxo1 in pre-B cells by counteracting PKB.
Figure 8: SLP-65 signaling regulates the PI(3)K-PKB-Foxo3a pathway in BCR+ cells.

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Acknowledgements

We thank C. Eschbach and I. Fiedler for technical assistance, A. Wuerch for cell sorting, P.J. Nielsen for critical discussion, B.A. Hemmings (Friedrich Miescher Institute for Biomedical Research) for the cDNA encoding a myristoylated PKB, W.S. Pear (University of Pennsylvania School of Medicine) for pMIG, B.M.T. Burgering (University Medical Center Utrecht) for a plasmid containing the Foxo3a-A3 construct, and R.Y. Tsien (University of California San Diego) for the tdTomato construct. Supported by the Claudia Adam Barr Foundation and the Damon Runyon Cancer Research Foundation (J.-H.P.), the Robert A. and Renee E. Belfer Foundation for Innovative Cancer Science (R.A.D.), the Deutsche Forschungsgemeinschaft (SFB620 and SFB746) and the Excellence Initiative of the German federal and state governments (EXC294).

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Authors and Affiliations

  1. Max Planck Institute for Immunobiology, 79108 Freiburg, Germany, and Centre for Biological Signalling Studies (BIOSS) at the Faculty of Biology, Albert-Ludwigs University of Freiburg, Freiburg, 79104, Germany

    Sebastian Herzog, Eva Hug, Sonja Meixlsperger, Michael Reth & Hassan Jumaa

  2. Departments of Medical Oncology, Center for Applied Cancer Science, Medicine and Genetics, Dana-Farber Cancer Institute, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Ji-Hye Paik & Ronald A DePinho

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  1. Sebastian Herzog
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Contributions

S.H. designed experiments, did all experimental studies unless otherwise indicated and wrote the manuscript. E.H. established the correlation between receptor expression and PKB regulation. S.M. established the inducible PKB (ERT2-PKB) system and performed the respective experiment. J.-H.P. and R.A.D. provided suggestions and mice deficient for Foxo3a. M.R. provided suggestions for experimental design. H.J. designed experiments, supervised the study, developed the concept and wrote the manuscript together with S.H.

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Correspondence to Hassan Jumaa.

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Herzog, S., Hug, E., Meixlsperger, S. et al. SLP-65 regulates immunoglobulin light chain gene recombination through the PI(3)K-PKB-Foxo pathway. Nat Immunol 9, 623–631 (2008). https://doi.org/10.1038/ni.1616

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