The "dispensable" portion of RAG2 is necessary for efficient V-to-DJ rearrangement during B and T cell development

Hong Erh Liang; Lih Yun Hsu; Dragana Cado; Lindsay G. Cowell; Garnett Kelsoe; Mark S. Schlissel

doi:10.1016/S1074-7613(02)00448-X

The "dispensable" portion of RAG2 is necessary for efficient V-to-DJ rearrangement during B and T cell development

Hong Erh Liang, Lih Yun Hsu, Dragana Cado, Lindsay G. Cowell, Garnett Kelsoe, Mark S. Schlissel

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

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Abstract

Previous in vitro studies defined the minimal regions of RAG1 and RAG2 essential for V(D)J recombination. In order to characterize the role of the C-terminal "dispensable" portion of RAG2, we generated core-RAG2 knock-in mice. We found that the core-RAG2-containing recombinase complex is selectively defective in catalyzing V-to-DJ rearrangement at the IgH and TCRβ loci, resulting in partial developmental blocks in B and T lymphopoiesis. Analysis of recombination intermediates showed defects at the cleavage phase of the reaction. We also observed a reduction in overall recombinase activity in core-RAG2-expressing thymocytes, leading us to suggest that the interaction of a defective recombinase with RSS sequences unique to VH and Vβ gene segments may underlie the specific V-to-DJ rearrangement defect in core-RAG2 mice.

Original language	English (US)
Pages (from-to)	639-651
Number of pages	13
Journal	Immunity
Volume	17
Issue number	5
DOIs	https://doi.org/10.1016/S1074-7613(02)00448-X
State	Published - Nov 1 2002

ASJC Scopus subject areas

Immunology and Allergy
Immunology
Infectious Diseases

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10.1016/S1074-7613(02)00448-X

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title = "The {"}dispensable{"} portion of RAG2 is necessary for efficient V-to-DJ rearrangement during B and T cell development",

abstract = "Previous in vitro studies defined the minimal regions of RAG1 and RAG2 essential for V(D)J recombination. In order to characterize the role of the C-terminal {"}dispensable{"} portion of RAG2, we generated core-RAG2 knock-in mice. We found that the core-RAG2-containing recombinase complex is selectively defective in catalyzing V-to-DJ rearrangement at the IgH and TCRβ loci, resulting in partial developmental blocks in B and T lymphopoiesis. Analysis of recombination intermediates showed defects at the cleavage phase of the reaction. We also observed a reduction in overall recombinase activity in core-RAG2-expressing thymocytes, leading us to suggest that the interaction of a defective recombinase with RSS sequences unique to VH and Vβ gene segments may underlie the specific V-to-DJ rearrangement defect in core-RAG2 mice.",

author = "Liang, {Hong Erh} and Hsu, {Lih Yun} and Dragana Cado and Cowell, {Lindsay G.} and Garnett Kelsoe and Schlissel, {Mark S.}",

note = "Funding Information: We thank Drs. Phil Leder and Fred Alt for the TC-1 ES cells, Dr. Yangsong Gu for ES cell technique, Dr. Hua Gu for pLZNeo vector, Drs. David Schatz, Alfred Lee, Ken Murphy, Bill Sha, and Naomi Rosenberg for the retroviral constructs, Dr. Stephen Desiderio for RAG2 antibody, and Mr. Hector Nolla for help with flow cytometry. This manuscript was improved by the helpful criticisms of various members of the Schlissel lab, several anonymous reviewers, and Drs. Astar Winoto and David Raulet. This work was funded by a grant from the NIH to M.S.S. (AI40227) and an Arthritis Foundation Biomedical Science Grant (M.S.S.).",

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AU - Liang, Hong Erh

AU - Hsu, Lih Yun

AU - Cado, Dragana

AU - Cowell, Lindsay G.

AU - Kelsoe, Garnett

AU - Schlissel, Mark S.

N1 - Funding Information: We thank Drs. Phil Leder and Fred Alt for the TC-1 ES cells, Dr. Yangsong Gu for ES cell technique, Dr. Hua Gu for pLZNeo vector, Drs. David Schatz, Alfred Lee, Ken Murphy, Bill Sha, and Naomi Rosenberg for the retroviral constructs, Dr. Stephen Desiderio for RAG2 antibody, and Mr. Hector Nolla for help with flow cytometry. This manuscript was improved by the helpful criticisms of various members of the Schlissel lab, several anonymous reviewers, and Drs. Astar Winoto and David Raulet. This work was funded by a grant from the NIH to M.S.S. (AI40227) and an Arthritis Foundation Biomedical Science Grant (M.S.S.).

PY - 2002/11/1

Y1 - 2002/11/1

N2 - Previous in vitro studies defined the minimal regions of RAG1 and RAG2 essential for V(D)J recombination. In order to characterize the role of the C-terminal "dispensable" portion of RAG2, we generated core-RAG2 knock-in mice. We found that the core-RAG2-containing recombinase complex is selectively defective in catalyzing V-to-DJ rearrangement at the IgH and TCRβ loci, resulting in partial developmental blocks in B and T lymphopoiesis. Analysis of recombination intermediates showed defects at the cleavage phase of the reaction. We also observed a reduction in overall recombinase activity in core-RAG2-expressing thymocytes, leading us to suggest that the interaction of a defective recombinase with RSS sequences unique to VH and Vβ gene segments may underlie the specific V-to-DJ rearrangement defect in core-RAG2 mice.

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The "dispensable" portion of RAG2 is necessary for efficient V-to-DJ rearrangement during B and T cell development

Abstract

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