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Clonal allelic predetermination of immunoglobulin-κ rearrangement

Nature volume 490, pages 561–565 (2012)Cite this article

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Abstract

Although most genes are expressed biallelically, a number of key genomic sites—including immune and olfactory receptor regions—are controlled monoallelically in a stochastic manner, with some cells expressing the maternal allele and others the paternal allele in the target tissue1,2. Very little is known about how this phenomenon is regulated and programmed during development. Here, using mouse immunoglobulin-κ (Igκ) as a model system, we demonstrate that although individual haematopoietic stem cells are characterized by allelic plasticity, early lymphoid lineage cells become committed to the choice of a single allele, and this decision is then stably maintained in a clonal manner that predetermines monoallelic rearrangement in B cells. This is accompanied at the molecular level by underlying allelic changes in asynchronous replication timing patterns at the κ locus. These experiments may serve to define a new concept of stem cell plasticity.

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Figure 1: Igκ allelic choice for rearrangement is predetermined and clonally inherited in pre-B cells.
Figure 2: Igκ-allele-specific ChIP analyses in pre-B cells.
Figure 3: Asynchronous Igκ replication.
Figure 4: Establishment of Igκ allelic choice.

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Acknowledgements

We thank M. Nussenzweig for providing the Igκm/h mice, A. Waisman for providing the IiMOG-marked ES cells, A. G. Fisher and V. Azuara for guiding us through the replication timing analysis by S-phase fractionation assay, M. Inlay for coaching us on the purification and culturing of MPPs and CLPs, A. Wutz for providing some of the single-cell clones, Y. Smith for image analysis and L. Dempsey for sharing ideas. This work was supported by research grants from the Israel Academy of Sciences (H.C., Y.B.), National Institutes of Health (Y.B.), the Israel Cancer Research Foundation (H.C., Y.B.), the European Community 5th Framework Quality of Life Program (Y.B.), Lew Sanders (H.C.) and Norton Herrick (H.C.).

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Author notes

  1. Maya Tevlin

    Present address: Present address: Laboratory of Developmental Genetics, Rockefeller University, 1230 York Avenue, New York, New York 10065, USA.,

  2. Marganit Farago, Chaggai Rosenbluh and Maya Tevlin: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Developmental Biology and Cancer Research, Institute for Medical Research Israel–Canada, Hebrew University Medical School, POB 12272, Ein Kerem, Jerusalem 91120, Israel,

    Marganit Farago, Chaggai Rosenbluh, Maya Tevlin, Shira Fraenkel, Sharon Schlesinger, Hagit Masika, Masha Gouzman, Howard Cedar & Yehudit Bergman

  2. Department of Immunobiology, Yale University School of Medicine, 300 Cedar Street, PO Box 208011, New Haven, Connecticut 06520, USA,

    Grace Teng & David Schatz

  3. Howard Hughes Medical Institute, Yale University School of Medicine, 300 Cedar Street, New Haven, Connecticut 06520, USA,

    David Schatz

  4. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, 76100, Israel

    Yoach Rais & Jacob H. Hanna

  5. Department of Immunology, Weizmann Institute of Science, Rehovot, 76100, Israel

    Alexander Mildner & Steffen Jung

  6. Department of Medicine, Center for Regenerative Medicine (CReM), Boston University School of Medicine, 650 Albany Street, Suite 513, Boston, Massachusetts 02118, USA,

    Gustavo Mostoslavsky

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Contributions

M.F. and C.R. designed the experiments, did the research, interpreted the results and assisted in writing the manuscript; M.T. initiated the allelic ChIP and DNA replication analyses; S.F. performed the allelic VJκ rearrangement in B6/Cast clones; S.S. studied asynchronous replication in ES cells; H.M. studied asynchronous replication in HSCs, MPPs and CLPs; G.T. and D.S. helped with Rag2 ChIP assays; Y.R. and J.H.H. generated the LN3 (Vβ1NT/+) mice from the appropriate ES cells; M.G., A.M. and S.J. designed and helped with the MPP and CLP reconstitution, and FACS experiments; G.M. designed and helped with the HSCs, pro-B, MPP and CLP reconstitution experiments; and H.C. and Y.B. directed the study and wrote the manuscript.

Corresponding authors

Correspondence to Howard Cedar or Yehudit Bergman.

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The authors declare no competing financial interests.

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Farago, M., Rosenbluh, C., Tevlin, M. et al. Clonal allelic predetermination of immunoglobulin-κ rearrangement. Nature 490, 561–565 (2012). https://doi.org/10.1038/nature11496

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