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Endocytosed BCRs sequentially regulate MAPK and Akt signaling pathways from intracellular compartments

Nature Immunology volume 12, pages 1119–1126 (2011)Cite this article

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A Corrigendum to this article was published on 19 January 2012

This article has been updated

Abstract

Binding of antigen to the B cell antigen receptor (BCR) triggers both BCR signaling and endocytosis. How endocytosis regulates BCR signaling remains unknown. Here we report that BCR signaling was not extinguished by endocytosis of BCRs; instead, BCR signaling initiated at the plasma membrane continued as the BCR trafficked intracellularly with the sequential phosphorylation of kinases. Blocking the endocytosis of BCRs resulted in the recruitment of both proximal and downstream kinases to the plasma membrane, where mitogen-activated protein kinases (MAPKs) were hyperphosphorylated and the kinase Akt and its downstream target Foxo were hypophosphorylated, which led to the dysregulation of gene transcription controlled by these pathways. Thus, the cellular location of the BCR serves to compartmentalize kinase activation to regulate the outcome of signaling.

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Figure 1: Spatial distribution of proximal and downstream phosphorylated kinases after BCR crosslinking.
Figure 2: Colocalization of proximal and downstream phosphorylated kinases with the BCR.
Figure 3: Colocalization of phosphorylated kinases with TfR and LAMP-1 after BCR crosslinking.
Figure 4: Temporal and spatial distribution of the phosphorylation of kinases in the BCR signaling pathway.
Figure 5: Dynasore blocks the internalization of BCRs.
Figure 6: The subcellular distribution of phosphorylated kinases in dynasore-treated B cells.
Figure 7: Inhibiting endocytosis of the BCR alters the amount of phosphorylation of kinases in the BCR signaling pathway.
Figure 8: Blocking endocytosis of the BCR blocks signals downstream of Akt and dysregulates transcription.

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Change history

  • 10 November 2011

    In the version of this article initially published, the authors refer to the phosphorylation of Lyn observed after crosslinking of the B cell antigen receptor (BCR) as indicating activation of Lyn. However, the antibody to phosphorylated Lyn used (Cell Signaling Technologies catalog number 2731) recognizes Lyn phosphorylated at Tyr507, the negative regulatory site, and thus recognizes the inactive form of Lyn. Antibodies for the detection of the active form of Lyn (phosphorylated at Tyr396) are not available at present. However, the authors conclude that after crosslinking of the BCR, active Lyn must be in dynamic equilibrium with inactive Lyn at BCR clusters, as active Lyn is required for the observed phosphorylation of downstream kinases. They state that the following overall conclusions of the paper remain unchanged: BCR signaling is initiated at the plasma membrane and continues as the BCR traffics into the cell with sequential phosphorylation of kinases, and endocytosis is essential to regulate the outcome of signal. Nonetheless, they would like to correct this so readers are not misled about the activation state of Lyn observed in their studies. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank T. Kirchhausen (Harvard Medical School) for dynasore. Supported by the Intramural Research Program of the US National Institutes of Health, National Institute of Allergy and Infectious Diseases.

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Authors and Affiliations

  1. Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA

    Akanksha Chaturvedi, Rebecca Martz, Michael Waisberg & Susan K Pierce

  2. Microscopy Unit, Rocky Mountain Laboratories, Research Technologies Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA

    David Dorward

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Contributions

A.C. designed and did the experiments, analyzed data and wrote the manuscript; R.M. did experiments and analyzed the data; D.D. did TEM and interpreted the images; M.W. designed transcription analysis and analyzed the quantitative PCR data; and S.K.P. designed experiments, analyzed data and wrote the manuscript.

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Correspondence to Akanksha Chaturvedi or Susan K Pierce.

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The authors declare no competing financial interests.

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Supplementary Figures 1–7 (PDF 9340 kb)

Supplementary Data Set

Ct values of complete data sets for Fig. 8b and 8c. (XLS 84 kb)

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Chaturvedi, A., Martz, R., Dorward, D. et al. Endocytosed BCRs sequentially regulate MAPK and Akt signaling pathways from intracellular compartments. Nat Immunol 12, 1119–1126 (2011). https://doi.org/10.1038/ni.2116

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