A non-B-DNA structure at the Bcl-2 major breakpoint region is cleaved by the RAG complex (original) (raw)

Nature volume 428, pages 88–93 (2004)Cite this article

Abstract

The causes of spontaneous chromosomal translocations in somatic cells of biological organisms are largely unknown, although double-strand DNA breaks are required in all proposed mechanisms1,2,3,4,5. The most common chromosomal abnormality in human cancer is the reciprocal translocation between chromosomes 14 and 18 (t(14;18)), which occurs in follicular lymphomas. The break at the immunoglobulin heavy-chain locus on chromosome 14 is an interruption of the normal V(D)J recombination process. But the breakage on chromosome 18, at the Bcl-2 gene, occurs within a confined 150-base-pair region (the major breakpoint region or Mbr) for reasons that have remained enigmatic. We have reproduced key features of the translocation process on an episome that propagates in human cells. The RAG complex—which is the normal enzyme for DNA cleavage at V, D or J segments—nicks the Bcl-2 Mbr in vitro and in vivo in a manner that reflects the pattern of the chromosomal translocations; however, the Mbr is not a V(D)J recombination signal. Rather the Bcl-2 Mbr assumes a non-B-form DNA structure within the chromosomes of human cells at 20–30% of alleles. Purified DNA assuming this structure contains stable regions of single-strandedness, which correspond well to the translocation regions in patients. Hence, a stable non-B-DNA structure in the human genome appears to be the basis for the fragility of the Bcl-2 Mbr, and the RAG complex is able to cleave this structure.

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Acknowledgements

We thank S. J. Korsmeyer for guidance during the early phases of this work. We also thank I. Haworth, J. S. Lee, P. Chastian and D. Shibata for discussions of the work, and NIH for grants to M.R.L.

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Authors and Affiliations

  1. Norris Comprehensive Cancer Center, Room 5428, University of Southern California Keck School of Medicine, 1441 Eastlake Ave., MC9176, Los Angeles, California, 90033, USA
    Sathees C. Raghavan, Xiantuo Wu, Chih-Lin Hsieh & Michael R. Lieber
  2. Departments of Pathology, University of Southern California Keck School of Medicine, 1441 Eastlake Ave., MC9176, Los Angeles, California, 90033, USA
    Sathees C. Raghavan & Michael R. Lieber
  3. Biochemistry & Molecular Biology, University of Southern California Keck School of Medicine, 1441 Eastlake Ave., MC9176, Los Angeles, California, 90033, USA
    Chih-Lin Hsieh & Michael R. Lieber
  4. Urology, University of Southern California Keck School of Medicine, 1441 Eastlake Ave., MC9176, Los Angeles, California, 90033, USA
    Chih-Lin Hsieh
  5. Biological Science and Molecular Microbiology & Immunology, University of Southern California Keck School of Medicine, 1441 Eastlake Ave., MC9176, Los Angeles, California, 90033, USA
    Michael R. Lieber
  6. Department of Medical Microbiology and Immunology, Creighton University School of Medicine, 2500 California Plaza, Omaha, Nebraska, 68178, USA
    Patrick C. Swanson

Authors

  1. Sathees C. Raghavan
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  2. Patrick C. Swanson
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  3. Xiantuo Wu
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  4. Chih-Lin Hsieh
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  5. Michael R. Lieber
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Correspondence toMichael R. Lieber.

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Raghavan, S., Swanson, P., Wu, X. et al. A non-B-DNA structure at the Bcl-2 major breakpoint region is cleaved by the RAG complex.Nature 428, 88–93 (2004). https://doi.org/10.1038/nature02355

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