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Enhancer control of local accessibility to V(D)J recombinase.

Publication ,  Journal Article
McMurry, MT; Hernandez-Munain, C; Lauzurica, P; Krangel, MS
Published in: Mol Cell Biol
August 1997

We have studied the role of transcriptional enhancers in providing recombination signal sequence (RSS) accessibility to V(D)J recombinase by examining mice carrying a transgenic human T-cell receptor (TCR) delta gene minilocus. This transgene is composed of unrearranged variable (Vdelta and Vdelta2), diversity (Ddelta3), joining (Jdelta1 and Jdelta3), and constant (Cdelta) gene segments. Previous data indicated that with the TCR delta enhancer (Edelta) present in the Jdelta3-Cdelta intron, V(D)J recombination proceeds stepwise, first V to D and then VD to J. With the enhancer deleted or mutated, V-to-D rearrangement is intact, but VD-to-J rearrangement is inhibited. We proposed that Edelta is necessary for J segment but not D segment accessibility and that J segment inaccessibility in the enhancerless minilocus resulted in the observed V(D)J recombination phenotype. In this study, we tested this notion by using ligation-mediated PCR to assess the formation of recombination-activating gene (RAG)-dependent double-strand breaks (DSBs) at RSSs 3' of Ddelta3 and 5' of Jdelta1. In five lines of mice carrying multicopy integrants of constructs that either lacked Edelta or carried an inactivated Edelta, the frequency of DSBs 5' of Jdelta1 was dramatically reduced relative to that in the wild type, whereas the frequency of DSBs 3' of Ddelta3 was unaffected. We interpret these results to indicate that Edelta is required for Jdelta1 but not Ddelta3 accessibility within the minilocus, and we conclude that enhancers regulate V(D)J recombination by providing local accessibility to the recombinase. cis-acting elements other than Edelta must maintain Ddelta3 in an accessible state in the absence of Edelta. The analysis of DSB formation in a single-copy minilocus integrant indicates that efficient DSB formation at the accessible RSS 3' of Ddelta3 requires an accessible partner RSS, arguing that RSS synapsis is required for DSB formation in chromosomal substrates in vivo.

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Published In

Mol Cell Biol

DOI

ISSN

0270-7306

Publication Date

August 1997

Volume

17

Issue

8

Start / End Page

4553 / 4561

Location

United States

Related Subject Headings

  • VDJ Recombinases
  • Transgenes
  • Recombination, Genetic
  • Receptors, Antigen, T-Cell, gamma-delta
  • Polymerase Chain Reaction
  • Mice, Transgenic
  • Mice
  • Humans
  • Genes, RAG-1
  • Enhancer Elements, Genetic
 

Citation

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McMurry, M. T., Hernandez-Munain, C., Lauzurica, P., & Krangel, M. S. (1997). Enhancer control of local accessibility to V(D)J recombinase. Mol Cell Biol, 17(8), 4553–4561. https://doi.org/10.1128/MCB.17.8.4553
McMurry, M. T., C. Hernandez-Munain, P. Lauzurica, and M. S. Krangel. “Enhancer control of local accessibility to V(D)J recombinase.Mol Cell Biol 17, no. 8 (August 1997): 4553–61. https://doi.org/10.1128/MCB.17.8.4553.
McMurry MT, Hernandez-Munain C, Lauzurica P, Krangel MS. Enhancer control of local accessibility to V(D)J recombinase. Mol Cell Biol. 1997 Aug;17(8):4553–61.
McMurry, M. T., et al. “Enhancer control of local accessibility to V(D)J recombinase.Mol Cell Biol, vol. 17, no. 8, Aug. 1997, pp. 4553–61. Pubmed, doi:10.1128/MCB.17.8.4553.
McMurry MT, Hernandez-Munain C, Lauzurica P, Krangel MS. Enhancer control of local accessibility to V(D)J recombinase. Mol Cell Biol. 1997 Aug;17(8):4553–4561.

Published In

Mol Cell Biol

DOI

ISSN

0270-7306

Publication Date

August 1997

Volume

17

Issue

8

Start / End Page

4553 / 4561

Location

United States

Related Subject Headings

  • VDJ Recombinases
  • Transgenes
  • Recombination, Genetic
  • Receptors, Antigen, T-Cell, gamma-delta
  • Polymerase Chain Reaction
  • Mice, Transgenic
  • Mice
  • Humans
  • Genes, RAG-1
  • Enhancer Elements, Genetic