RAG1 and RAG2 cooperate in specific binding to the recombination signal sequence in vitro

Xianming Mo, Tu Bailin, Moshe J. Sadofsky

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

An essential step in the development of the vertebrate immune system is the DNA level rearrangement of the antigen receptor genes. This process, termed 'V(D)J recombination,' begins with DNA cleavage at the appropriate sites mediated by the two proteins RAG1 and RAG2. We report here that the two proteins cooperate to bind DNA with significantly higher specificity than either protein alone. Gel purification of the triple complex is performed in the absence of any cross-linking agents. Both proteins remain present in the complex, and UV cross-linking using iodouridine-containing probes shows that RAG1 makes close contacts in both the heptamer and nonamer motifs. The two proteins are also shown to associate with each other in the absence of any DNA. These findings refine our understanding of the protein-DNA interactions that accompany cleavage at the recombination signals.

Original languageEnglish (US)
Pages (from-to)7025-7031
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number11
DOIs
StatePublished - Mar 12 1999
Externally publishedYes

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Protein Sorting Signals
Genetic Recombination
DNA
Proteins
V(D)J Recombination
DNA Cleavage
Antigen Receptors
Gene Rearrangement
Immune system
Purification
Vertebrates
In Vitro Techniques
Immune System
Genes
Gels

ASJC Scopus subject areas

  • Biochemistry

Cite this

RAG1 and RAG2 cooperate in specific binding to the recombination signal sequence in vitro. / Mo, Xianming; Bailin, Tu; Sadofsky, Moshe J.

In: Journal of Biological Chemistry, Vol. 274, No. 11, 12.03.1999, p. 7025-7031.

Research output: Contribution to journalArticle

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