Abstract
Developing B cells express distinct classes of B cell antigen receptors (BCRs) that differ in their heavy chain (HC). Although only μHC is expressed in early stages, δHC-containing BCRs dominate on the surface of mature B cells. The reason for the tightly regulated expression of these receptors is poorly understood. Here we show that μHC was specifically required for precursor BCR (pre-BCR) function and that δHC was unable to form a functional pre-BCR. A conserved asparagine (N)-linked glycosylation site at position 46 (N46) in the first conserved domain of μHC was absolutely required for pre-BCR function, and swapping that domain with δHC resulted in a functional δHC-containing pre-BCR. When tested in the context of the BCR, μHC with a mutant N46 showed normal function, which indicated that N46-glycosylation is specifically required for pre-BCR function. Our results suggest an unexpected mode of pre-BCR function, in which binding of the surrogate light chain to N46 mediates autonomous crosslinking and, concomitantly, receptor formation.
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Acknowledgements
We thank R.Y. Tsien (University of California) for tdTomato; A.G. Rolink (University of Basel) for phycoerythrin-biotin–anti-λ5; J.C. Cambier (National Jewish Medical and Research Center and University of Colorado Health Sciences Center) for anti-immunoglobulin-α; P.J. Nielsen and F. Köhler for reading the manuscript and discussions; A. Wuerch and S. Hobitz for help with cell sorting; and U. Stauffer, N. Joswig and C. Johner for mouse work. Supported by Deutsche Forschungsgemeinschaft (SFB746 and SFB 620) and the Excellence Initiative of the German Federal and State Governments (GSC-4, Spemann Graduate School).
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R.Ü. and M.P.B. did experiments, analyzed data and contributed to the writing of the manuscript; C.E. did experiments and provided advice; T.W. analyzed the μHC-deficient mice; M.R. provided suggestions for experimental design; and H.J. supervised the work, designed experiments and wrote the manuscript.
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Übelhart, R., Bach, M., Eschbach, C. et al. N-linked glycosylation selectively regulates autonomous precursor BCR function. Nat Immunol 11, 759–765 (2010). https://doi.org/10.1038/ni.1903
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DOI: https://doi.org/10.1038/ni.1903
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