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Foxo1 directly regulates the transcription of recombination-activating genes during B cell development

Nature Immunology volume 9, pages 613–622 (2008)Cite this article

Abstract

Regulated expression of the recombinase RAG-1 and RAG-2 proteins is necessary for generating the vast repertoire of antigen receptors essential for adaptive immunity. Here, a retroviral cDNA library screen showed that the stress-regulated protein GADD45a activated transcription of the genes encoding RAG-1 and RAG-2 in transformed pro–B cells by a pathway requiring the transcription factor Foxo1. Foxo1 directly activated transcription of the Rag1-Rag2 locus throughout early B cell development, and a decrease in Foxo1 protein diminished the induction of Rag1 and Rag2 transcription in a model of receptor editing. We also found that transcription of Rag1 and Rag2 was repressed at the pro–B cell and immature B cell stages by the kinase Akt through its 'antagonism' of Foxo1 function. Thus, Foxo1 is a key regulator of Rag1 and Rag2 transcription in primary B cells.

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Figure 1: Identification of Gadd45a by a retroviral cDNA library screen for factors that induce Rag1 transcription in AMuLV-transformed B cells.
Figure 2: GADD45a induces Rag1 transcription by a MEKK4- and p38-dependent pathway.
Figure 3: Foxo1 functions 'downstream' of GADD45a to increase Rag1 transcription in AMuLV-transformed pro–B cells.
Figure 4: Overexpression of Foxo1 increases Rag1 transcription in primary B lymphocytes, whereas constitutively active Akt suppresses it.
Figure 5: A decrease in Foxo1 protein in primary B lymphocytes decreases Rag1 transcription at all stages of B cell development.
Figure 6: IL-7R and BCR signaling repress Rag transcription through the PI(3)K-Akt signaling axis.
Figure 7: A decrease in Foxo1 protein in primary immature B cells interferes with Rag1 and Rag2 transcript induction in response to BCR crosslinking.
Figure 8: Foxo1 acts directly on the Rag locus.

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Acknowledgements

We thank all who provided mice (N. Sakaguchi; Kumamoto University) and plasmid constructs (K. Murphy (University of Washington at St. Louis), A. Rao (Harvard Medical School), E. Olson (University of Texas Southwestern Medical Center), D. Fruman (University of California at Irvine), W. Sha (University of California at Berkeley), N. Rosenberg (Tufts Medical School) and G. Barton (University of California at Berkeley)); H. Nolla (Cancer Research Laboratories, University of California at Berkeley) for help with flow cytometry cell sorting; and P. Herzmark (University of California at Berkeley) for help and advice with live microscopy. Supported by the National Institutes of Health (RO1 HL48702 and RO1 AI57487 to M.S.S.).

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  1. Department of Molecular & Cell Biology, University of California at Berkeley, Berkeley, 94720, California, USA

    Rupesh H Amin & Mark S Schlissel

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  1. Rupesh H Amin
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R.H.A. did all the experiments; and R.H.A. and M.S.S. jointly designed the experiments and wrote the manuscript.

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Correspondence to Mark S Schlissel.

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Amin, R., Schlissel, M. Foxo1 directly regulates the transcription of recombination-activating genes during B cell development. Nat Immunol 9, 613–622 (2008). https://doi.org/10.1038/ni.1612

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