A non-B-DNA structure at the Bcl-2 major breakpoint region is cleaved by the RAG complex.
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 mechanisms. 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.
Raghavan
Sathees C
SC
Norris Comprehensive Cancer Center, Room 5428, University of Southern California Keck School of Medicine, 1441 Eastlake Ave., MC9176, Los Angeles, California 90033, USA.
Swanson
Patrick C
PC
Wu
Xiantuo
X
Hsieh
Chih-Lin
CL
Lieber
Michael R
MR
D000818:Animals; D002460:Cell Line; D019457:Chromosome Breakage / Q000235:genetics*; D002883:Chromosomes, Human, Pair 14 / Q000235:genetics; D002887:Chromosomes, Human, Pair 18 / Q000235:genetics; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D019254:Genes, bcl-2 / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D009687:Nuclear Proteins; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics; D013447:Sulfites / Q000378:metabolism; D014178:Translocation, Genetic / Q000235:genetics
D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D046911:Macromolecular Substances; D009687:Nuclear Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; C499311:RAG2 protein, human; C499308:Rag2 protein, mouse; D013447:Sulfites; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein; D004247:DNA; C042345:hydrogen sulfite