Footprint analysis of recombination signal sequences in the 12/23 synaptic complex of V(D)J recombination.
Mol Cell Biol
; 22(20): 7217-25, 2002 Oct.
Article
in En
| MEDLINE
| ID: mdl-12242298
ABSTRACT
In V(D)J joining of antigen receptor genes, two recombination signal sequences (RSSs), 12-RSS and 23-RSS, are paired and complexed with the protein products of recombination-activating genes RAG1 and RAG2. Using magnetic beads, we purified the pre- and postcleavage complexes of V(D)J joining and analyzed them by DNase I footprinting. In the precleavage synaptic complex, strong protection was seen not only in the 9-mer and spacer regions but also near the coding border of the 7-mer. This is a sharp contrast to the single RSS-RAG complex where the 9-mer plays a major role in the interaction. We also analyzed the postcleavage signal end complex by footprinting. Unlike what was seen with the precleavage complex, the entire 7-mer and its neighboring spacer regions were protected. The present study indicates that the RAG-RSS interaction in the 7-mer region drastically changes once the synaptic complex is formed for cleavage.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Protein Sorting Signals
/
DNA, Intergenic
/
DNA Nucleotidyltransferases
Language:
En
Journal:
Mol Cell Biol
Year:
2002
Document type:
Article
Affiliation country: