Double-strand break formation by the RAG complex at the Bcl-2 major breakpoint region and at other non-B DNA structures in vitro

Sathees C. Raghavan, Patrick Swanson, Yunmei Ma, Michael R. Lieber

Research output: Contribution to journalArticle

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Abstract

The most common chromosomal translocation in cancer, t(14;18) at the 150-bp bcl-2 major breakpoint region (Mbr), occurs in follicular lymphomas. The bcl-2 Mbr assumes a non-B DNA conformation, thus explaining its distinctive fragility. This non-B DNA structure is a target of the RAG complex in vivo, but not because of its primary sequence. Here we report that the RAG complex generates at least two independent nicks that lead to double-strand breaks in vitro, and this requires the non-B DNA structure at the bcl-2 Mbr. A 3-bp mutation is capable of abolishing the non-B structure formation and the double-strand breaks. The observations on the bcl-2 Mbr reflect more general properties of the RAG complex, which can bind and nick at duplex-single-strand transitions of other non-B DNA structures, resulting in double-strand breaks in vitro. Hence, the present study reveals novel insight into a third mechanism of action of RAGs on DNA, besides the standard heptamer/nonamer-mediated cleavage in V(D)J recombination and the in vitro transposase activity.

Original languageEnglish
Pages (from-to)5904-5919
Number of pages16
JournalMolecular and Cellular Biology
Volume25
Issue number14
DOIs
StatePublished - Jul 2005

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DNA
V(D)J Recombination
Transposases
Nucleic Acid Conformation
Genetic Translocation
Follicular Lymphoma
Mutation
In Vitro Techniques
Neoplasms

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Double-strand break formation by the RAG complex at the Bcl-2 major breakpoint region and at other non-B DNA structures in vitro. / Raghavan, Sathees C.; Swanson, Patrick; Ma, Yunmei; Lieber, Michael R.

In: Molecular and Cellular Biology, Vol. 25, No. 14, 07.2005, p. 5904-5919.

Research output: Contribution to journalArticle

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