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 language | English |
|---|---|
| Pages (from-to) | 5904-5919 |
| Number of pages | 16 |
| Journal | Molecular and Cellular Biology |
| Volume | 25 |
| Issue number | 14 |
| DOIs | |
| State | Published - Jul 2005 |
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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 journal › Article
}
TY - JOUR
T1 - Double-strand break formation by the RAG complex at the Bcl-2 major breakpoint region and at other non-B DNA structures in vitro
AU - Raghavan, Sathees C.
AU - Swanson, Patrick
AU - Ma, Yunmei
AU - Lieber, Michael R.
PY - 2005/7
Y1 - 2005/7
N2 - 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.
AB - 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.
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U2 - 10.1128/MCB.25.14.5904-5919.2005
DO - 10.1128/MCB.25.14.5904-5919.2005
M3 - Article
VL - 25
SP - 5904
EP - 5919
JO - Molecular and Cellular Biology
JF - Molecular and Cellular Biology
SN - 0270-7306
IS - 14
ER -