The scid recombination-inducible cell line: A model to study DNA-PK-independent V(D)J recombination

Matthew L. Brown, Sandra Lew, Yung Chang

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

To investigate the molecular mechanisms of the variable (diversity) joining (V(D)J) recombination process at an endogenous gene locus, recombination-inducible cell lines were made from both bcl-2-bearing severe combined immune deficiency (scid) homozygous and scid heterozyous (s/+) mice by transforming pre-B cells with the temperature-sensitive Abelson murine leukemia virus (ts-Ab-MLV). These transformants can be induced to undergo immunoglobulin light-chain gene rearrangements by incubating them at the non-permissive temperature. In the case of transformed scid cells, a significant amount of hairpin coding ends are accumulated during recombination induction, but few coding joints are generated. After being shifted to the permissive temperature, however, these cells are capable of opening hairpin ends and forming coding joints. Thus, ts-Ab-MLV transformed scid cells can be readily manipulated for both recombination cleavage and end resolution. However, unlike the rapid coding joint formation in s/+ cells that have the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs), the process for resolving coding ends in scid cells is slow and error prone, and also appears to be correlated with a reduction in the RAG1/2 expression. Apparently, this process is mediated by a DNA-PK-independent pathway. The fact that the activity of this pathway can be manipulated in vitro makes it possible to delineate the mechanisms in end opening, processing and joining. Therefore, these ts-Ab-MLV transformed scid cell lines offer a model to study the molecular nature as well as the regulation of the DNA-PK-independent pathway in coding end resolution.

Original languageEnglish (US)
Pages (from-to)21-26
Number of pages6
JournalImmunology Letters
Volume75
Issue number1
DOIs
StatePublished - Dec 1 2000

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Keywords

  • DNA-dependent protein kinase
  • RAG
  • Scid
  • V(D)J recombination

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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