Extrachromosomal recombination substrates recapitulate beyond 12/23 restricted V(D)J recombination in nonlymphoid cells

David Jung; Craig H. Bassing; Sebastian D. Fugmann; Hwei Ling Cheng; David G. Schatz; Frederick W. Alt

doi:10.1016/S1074-7613(02)00507-1

Extrachromosomal recombination substrates recapitulate beyond 12/23 restricted V(D)J recombination in nonlymphoid cells

David Jung, Craig H. Bassing, Sebastian D. Fugmann, Hwei Ling Cheng, David G. Schatz, Frederick W. Alt^*

^*Corresponding author for this work

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

59 Scopus citations

Abstract

V(D)J recombination occurs efficiently only between gene segments flanked by recombination signals (RSs) containing 12 and 23 base pair spacers (the 12/23 rule). A further limitation "beyond the 12/23 rule" (B12/23) exists at the TCRβ locus and ensures Dβ usage. Herein, we show that extrachromosomal V(D)J recombination substrates recapitulate B12/23 restriction in nonlymphoid cells. We further demonstrate that the Vβ coding flank, the 12-RS heptamer/nonamer, and the 23-RS spacer each can significantly influence B12/23 restriction. Finally, purified core RAG1 and RAG2 proteins (together with HMG2) also reproduce B12/23 restriction in a cell-free system. Our findings indicate that B12/23 restriction of V(D)J recombination is cemented at the level of interactions between the RAG proteins and TCRβ RS sequences.

Original language	English
Pages (from-to)	65-74
Number of pages	10
Journal	Immunity
Volume	18
Issue number	1
DOIs	https://doi.org/10.1016/S1074-7613(02)00507-1
State	Published - 01 01 2003
Externally published	Yes

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title = "Extrachromosomal recombination substrates recapitulate beyond 12/23 restricted V(D)J recombination in nonlymphoid cells",

abstract = "V(D)J recombination occurs efficiently only between gene segments flanked by recombination signals (RSs) containing 12 and 23 base pair spacers (the 12/23 rule). A further limitation {"}beyond the 12/23 rule{"} (B12/23) exists at the TCRβ locus and ensures Dβ usage. Herein, we show that extrachromosomal V(D)J recombination substrates recapitulate B12/23 restriction in nonlymphoid cells. We further demonstrate that the Vβ coding flank, the 12-RS heptamer/nonamer, and the 23-RS spacer each can significantly influence B12/23 restriction. Finally, purified core RAG1 and RAG2 proteins (together with HMG2) also reproduce B12/23 restriction in a cell-free system. Our findings indicate that B12/23 restriction of V(D)J recombination is cemented at the level of interactions between the RAG proteins and TCRβ RS sequences.",

author = "David Jung and Bassing, {Craig H.} and Fugmann, {Sebastian D.} and Cheng, {Hwei Ling} and Schatz, {David G.} and Alt, {Frederick W.}",

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T1 - Extrachromosomal recombination substrates recapitulate beyond 12/23 restricted V(D)J recombination in nonlymphoid cells

AU - Jung, David

AU - Bassing, Craig H.

AU - Fugmann, Sebastian D.

AU - Cheng, Hwei Ling

AU - Schatz, David G.

AU - Alt, Frederick W.

PY - 2003/1/1

Y1 - 2003/1/1

N2 - V(D)J recombination occurs efficiently only between gene segments flanked by recombination signals (RSs) containing 12 and 23 base pair spacers (the 12/23 rule). A further limitation "beyond the 12/23 rule" (B12/23) exists at the TCRβ locus and ensures Dβ usage. Herein, we show that extrachromosomal V(D)J recombination substrates recapitulate B12/23 restriction in nonlymphoid cells. We further demonstrate that the Vβ coding flank, the 12-RS heptamer/nonamer, and the 23-RS spacer each can significantly influence B12/23 restriction. Finally, purified core RAG1 and RAG2 proteins (together with HMG2) also reproduce B12/23 restriction in a cell-free system. Our findings indicate that B12/23 restriction of V(D)J recombination is cemented at the level of interactions between the RAG proteins and TCRβ RS sequences.

AB - V(D)J recombination occurs efficiently only between gene segments flanked by recombination signals (RSs) containing 12 and 23 base pair spacers (the 12/23 rule). A further limitation "beyond the 12/23 rule" (B12/23) exists at the TCRβ locus and ensures Dβ usage. Herein, we show that extrachromosomal V(D)J recombination substrates recapitulate B12/23 restriction in nonlymphoid cells. We further demonstrate that the Vβ coding flank, the 12-RS heptamer/nonamer, and the 23-RS spacer each can significantly influence B12/23 restriction. Finally, purified core RAG1 and RAG2 proteins (together with HMG2) also reproduce B12/23 restriction in a cell-free system. Our findings indicate that B12/23 restriction of V(D)J recombination is cemented at the level of interactions between the RAG proteins and TCRβ RS sequences.

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DO - 10.1016/S1074-7613(02)00507-1

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AN - SCOPUS:0037242580

SN - 1074-7613

VL - 18

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EP - 74

JO - Immunity

JF - Immunity

IS - 1

ER -

Extrachromosomal recombination substrates recapitulate beyond 12/23 restricted V(D)J recombination in nonlymphoid cells

Abstract

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