Hairpin coding end opening is mediated by RAG1 and RAG2 proteins

Eva Besmer; Jorge Mansilla-Soto; Sylvanie Cassard; Dennis J. Sawchuk; Greg Brown; Moshe Sadofsky; Susanna M. Lewis; Michel C. Nussenzweig; Patricia Cortes

doi:10.1016/S1097-2765(00)80296-8

Hairpin coding end opening is mediated by RAG1 and RAG2 proteins

Eva Besmer, Jorge Mansilla-Soto, Sylvanie Cassard, Dennis J. Sawchuk, Greg Brown, Moshe Sadofsky, Susanna M. Lewis, Michel C. Nussenzweig, Patricia Cortes

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

115 Scopus citations

Abstract

Despite the importance of hairpin opening in antigen receptor gene assembly, the molecular machinery that mediates this reaction has not been defined. Here, we show that RAG1 plus RAG2 can open DNA hairpins. Hairpin opening by RAGs is not sequence specific, but in Mg²⁺, hairpin opening occurs only in the context of a regulated cleavage complex. The chemical mechanism of harpin opening by RAGs resembles RSS cleavage and 3′ end processing by HIV integrase and Mu transposase in that these reactions can proceed through alcoholysis. Mutations in either RAG1 or RAG2 that interfere with RSS cleavage also interfere with hairpin opening, suggesting that RAGs have a single active site that catalyzes several distinct DNA cleavage reactions.

Original language	English (US)
Pages (from-to)	817-828
Number of pages	12
Journal	Molecular Cell
Volume	2
Issue number	6
DOIs	https://doi.org/10.1016/S1097-2765(00)80296-8
State	Published - Dec 1998
Externally published	Yes

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1016/S1097-2765(00)80296-8

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title = "Hairpin coding end opening is mediated by RAG1 and RAG2 proteins",

abstract = "Despite the importance of hairpin opening in antigen receptor gene assembly, the molecular machinery that mediates this reaction has not been defined. Here, we show that RAG1 plus RAG2 can open DNA hairpins. Hairpin opening by RAGs is not sequence specific, but in Mg2+, hairpin opening occurs only in the context of a regulated cleavage complex. The chemical mechanism of harpin opening by RAGs resembles RSS cleavage and 3′ end processing by HIV integrase and Mu transposase in that these reactions can proceed through alcoholysis. Mutations in either RAG1 or RAG2 that interfere with RSS cleavage also interfere with hairpin opening, suggesting that RAGs have a single active site that catalyzes several distinct DNA cleavage reactions.",

author = "Eva Besmer and Jorge Mansilla-Soto and Sylvanie Cassard and Sawchuk, {Dennis J.} and Greg Brown and Moshe Sadofsky and Lewis, {Susanna M.} and Nussenzweig, {Michel C.} and Patricia Cortes",

note = "Funding Information: We thank Juan C{\'a}rcamo for comments on the manuscript and very helpful discussions. D. J. S. is supported by a Natural Sciences and Engineering Research Council of Canada (NSERC) PGS-B fellowship. S. M. L. is supported by a grant from the Medical Research Council of Canada and is a Scientist of the National Cancer Institute of Canada. M. C. N. is supported by the National Institutes of Health and is an associate investigator of the Howard Hughes Medical Institute. This work was supported in part by a grant from the Arthritis Foundation to P. C.",

year = "1998",

month = dec,

doi = "10.1016/S1097-2765(00)80296-8",

language = "English (US)",

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journal = "Molecular Cell",

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T1 - Hairpin coding end opening is mediated by RAG1 and RAG2 proteins

AU - Besmer, Eva

AU - Mansilla-Soto, Jorge

AU - Cassard, Sylvanie

AU - Sawchuk, Dennis J.

AU - Brown, Greg

AU - Sadofsky, Moshe

AU - Lewis, Susanna M.

AU - Nussenzweig, Michel C.

AU - Cortes, Patricia

N1 - Funding Information: We thank Juan Cárcamo for comments on the manuscript and very helpful discussions. D. J. S. is supported by a Natural Sciences and Engineering Research Council of Canada (NSERC) PGS-B fellowship. S. M. L. is supported by a grant from the Medical Research Council of Canada and is a Scientist of the National Cancer Institute of Canada. M. C. N. is supported by the National Institutes of Health and is an associate investigator of the Howard Hughes Medical Institute. This work was supported in part by a grant from the Arthritis Foundation to P. C.

PY - 1998/12

Y1 - 1998/12

N2 - Despite the importance of hairpin opening in antigen receptor gene assembly, the molecular machinery that mediates this reaction has not been defined. Here, we show that RAG1 plus RAG2 can open DNA hairpins. Hairpin opening by RAGs is not sequence specific, but in Mg2+, hairpin opening occurs only in the context of a regulated cleavage complex. The chemical mechanism of harpin opening by RAGs resembles RSS cleavage and 3′ end processing by HIV integrase and Mu transposase in that these reactions can proceed through alcoholysis. Mutations in either RAG1 or RAG2 that interfere with RSS cleavage also interfere with hairpin opening, suggesting that RAGs have a single active site that catalyzes several distinct DNA cleavage reactions.

AB - Despite the importance of hairpin opening in antigen receptor gene assembly, the molecular machinery that mediates this reaction has not been defined. Here, we show that RAG1 plus RAG2 can open DNA hairpins. Hairpin opening by RAGs is not sequence specific, but in Mg2+, hairpin opening occurs only in the context of a regulated cleavage complex. The chemical mechanism of harpin opening by RAGs resembles RSS cleavage and 3′ end processing by HIV integrase and Mu transposase in that these reactions can proceed through alcoholysis. Mutations in either RAG1 or RAG2 that interfere with RSS cleavage also interfere with hairpin opening, suggesting that RAGs have a single active site that catalyzes several distinct DNA cleavage reactions.

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Hairpin coding end opening is mediated by RAG1 and RAG2 proteins

Abstract

ASJC Scopus subject areas

UN SDGs

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