Abstract
In V(D)J joining of antigen receptor genes, two recombination signal sequences (RSSs), 12- and 23-RSSs, form a complex with the protein products of recombination activating genes, RAG1 and RAG2. DNaseI footprinting demonstrates that the interaction of RAG proteins with substrate RSS DNA is not just limited to the signal region but involves the coding sequence as well. Joining mutants of RAG1 and RAG2 demonstrate impaired interactions with the coding region in both pre- and postcleavage type complexes. A possible role of this RAG coding region interaction is discussed in the context of V(D)J recombination.
Highlights
V(D)J recombination plays key roles in activating and diversifying the antigen receptor genes in lymphocytes [1]
To prevent the double strand cleavage of nicked substrates, the complex was formed in the presence of Ca2ϩ, and the 3Ј-OH at the nick was reduced to the 3Ј-deoxy form
We have demonstrated that recombination signal sequences (RSSs) DNA is protected in the signal region and in the adjacent coding region in the synaptic complex
Summary
V(D)J recombination plays key roles in activating and diversifying the antigen receptor genes in lymphocytes [1]. DNaseI footprinting demonstrates that the interaction of RAG proteins with substrate RSS DNA is not just limited to the signal region but involves the coding sequence as well. We have found that the RAG proteins interact with the RSS DNA in the signal region, and in the coding region in the both pre-cleavage and post-cleavage type synaptic complexes.
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