Initiation of V(D)J recombination in a cell-free system

Dik C. van Gent; J. Fraser McBlane; Dale A. Ramsden; Moshe J. Sadofsky; Joanne E. Hesse; Martin Gellert

doi:10.1016/0092-8674(95)90012-8

Initiation of V(D)J recombination in a cell-free system

Dik C. van Gent^*, J. Fraser McBlane, Dale A. Ramsden, Moshe J. Sadofsky, Joanne E. Hesse, Martin Gellert

^*Corresponding author for this work

National Institutes of Health

Research output: Contribution to journal › Article › Academic › peer-review

274 Citations (Scopus)

Abstract

Cells performing V(D)J recombination make specific cuts in DNA at recombination signal sequences. Here, we show that nuclear extracts of pre-B cell lines carry out this specific cleavage. The products of cleavage are the same as found previously in thymocytes: fulllength, blunt, 5′-phosphorylated signal ends, and covalently sealed (hairpin) coding ends. A complete signal sequence is required. Recombinant RAG1 protein greatly increases activity and complements an inactive extract from a RAG1 (-/-) pre-B cell line. When the extracts are fractionated, cleavage activity correlates with the presence of RAG2 protein. These results suggest that RAG1 and RAG2 are components of the V(D)J recombinase.

Original language	English
Pages (from-to)	925-934
Number of pages	10
Journal	Cell
Volume	81
Issue number	6
DOIs	https://doi.org/10.1016/0092-8674(95)90012-8
Publication status	Published - 16 Jun 1995
Externally published	Yes

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title = "Initiation of V(D)J recombination in a cell-free system",

abstract = "Cells performing V(D)J recombination make specific cuts in DNA at recombination signal sequences. Here, we show that nuclear extracts of pre-B cell lines carry out this specific cleavage. The products of cleavage are the same as found previously in thymocytes: fulllength, blunt, 5′-phosphorylated signal ends, and covalently sealed (hairpin) coding ends. A complete signal sequence is required. Recombinant RAG1 protein greatly increases activity and complements an inactive extract from a RAG1 (-/-) pre-B cell line. When the extracts are fractionated, cleavage activity correlates with the presence of RAG2 protein. These results suggest that RAG1 and RAG2 are components of the V(D)J recombinase.",

author = "\{van Gent\}, \{Dik C.\} and \{Fraser McBlane\}, J. and Ramsden, \{Dale A.\} and Sadofsky, \{Moshe J.\} and Hesse, \{Joanne E.\} and Martin Gellert",

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T1 - Initiation of V(D)J recombination in a cell-free system

AU - van Gent, Dik C.

AU - Fraser McBlane, J.

AU - Ramsden, Dale A.

AU - Sadofsky, Moshe J.

AU - Hesse, Joanne E.

AU - Gellert, Martin

PY - 1995/6/16

Y1 - 1995/6/16

N2 - Cells performing V(D)J recombination make specific cuts in DNA at recombination signal sequences. Here, we show that nuclear extracts of pre-B cell lines carry out this specific cleavage. The products of cleavage are the same as found previously in thymocytes: fulllength, blunt, 5′-phosphorylated signal ends, and covalently sealed (hairpin) coding ends. A complete signal sequence is required. Recombinant RAG1 protein greatly increases activity and complements an inactive extract from a RAG1 (-/-) pre-B cell line. When the extracts are fractionated, cleavage activity correlates with the presence of RAG2 protein. These results suggest that RAG1 and RAG2 are components of the V(D)J recombinase.

AB - Cells performing V(D)J recombination make specific cuts in DNA at recombination signal sequences. Here, we show that nuclear extracts of pre-B cell lines carry out this specific cleavage. The products of cleavage are the same as found previously in thymocytes: fulllength, blunt, 5′-phosphorylated signal ends, and covalently sealed (hairpin) coding ends. A complete signal sequence is required. Recombinant RAG1 protein greatly increases activity and complements an inactive extract from a RAG1 (-/-) pre-B cell line. When the extracts are fractionated, cleavage activity correlates with the presence of RAG2 protein. These results suggest that RAG1 and RAG2 are components of the V(D)J recombinase.

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DO - 10.1016/0092-8674(95)90012-8

M3 - Article

C2 - 7781069

AN - SCOPUS:0028979702

SN - 0092-8674

VL - 81

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EP - 934

JO - Cell

JF - Cell

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ER -

Initiation of V(D)J recombination in a cell-free system

Abstract

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