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Essential roles of the κ light chain intronic enhancer and 3′ enhancer in κ rearrangement and demethylation

Nature Immunology volume 3, pages 463–468 (2002)Cite this article

Abstract

The κ intronic (MiEκ) and 3′ (3′Eκ) enhancers are both quantitatively important to, but not essential for, immunoglobulin κ rearrangement. To determine the functional redundancy between these two enhancers, B cells derived from mutant embryonic stem cells—in which both MiEκ and 3′Eκ were deleted on both κ alleles—were analyzed for κ rearrangement. Our findings indicate that these double-mutant B cells have essentially no κ rearrangement but do rearrange and express λ. Therefore, these two κ enhancers share essential roles in activating VκJκ rearrangement. Our findings also indicate that the two κ enhancers play overlapping and distinct roles in the demethylation of κ in B cells.

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Figure 1: Generation of EDK ES cells.
Figure 2: Analysis of serum κ and λ concentration by ELISA.
Figure 3: Flow cytometric analysis of κ+ and λ+ B cells.
Figure 4: Analysis of VκJκ rearrangement.
Figure 5: Southern blot analysis of κ-producing hybridomas derived from 3′Eκ−/− mice.
Figure 6: Analysis of κ germline transcripts in wild-type and mutant pre-B cells.
Figure 7: Analysis of the methylation status of the κ locus in wild-type and mutant splenic B cells by the bisulfite method.

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Acknowledgements

We thank M. Schlissel for 3′Eκ−/− mice and members of Alt laboratory for critical reading of this manuscript. Supported by NIH grant AI44838 (to Y. X.).

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Authors and Affiliations

  1. Division of Biology, Section of Molecular Biology, University of California, San Diego, 9500 Gilman Drive, La Jolla, 92093-0322, CA, USA

    Matthew Inlay & Yang Xu

  2. The Center for Blood Research, The Children's Hospital, Howard Hughes Medical Institute, Harvard Medical School, Boston, 02115, MA, USA

    Frederick W. Alt

  3. Division of Biology, California Institute of Technology, Pasadena, 91125, CA, USA

    David Baltimore

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Correspondence to Yang Xu.

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Inlay, M., Alt, F., Baltimore, D. et al. Essential roles of the κ light chain intronic enhancer and 3′ enhancer in κ rearrangement and demethylation. Nat Immunol 3, 463–468 (2002). https://doi.org/10.1038/ni790

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