Abstract
Maintenance of genomic integrity during antigen receptor gene rearrangements requires (1) regulated access of the V(D)J recombinase to specific loci and (2) generation of double-strand DNA breaks only after recognition of a pair of matched recombination signal sequences (RSSs). Here we recapitulate both key aspects of regulated recombinase accessibility in a cell-free system using plasmid substrates assembled into chromatin. We show that recruitment of the SWI/SNF chromatin-remodeling complex to both RSSs increases coupled cleavage by RAG1 and RAG2 proteins. SWI/SNF functions by altering local chromatin structure in the absence of RNA polymerase II-dependent transcription or histone modifications. These observations demonstrate a direct role for cis-sequence-regulated local chromatin remodeling in RAG1/2-dependent initiation of V(D)J recombination.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, N.I.H., Intramural
MeSH terms
-
Animals
-
Base Sequence*
-
Chromatin / genetics
-
Chromatin / metabolism
-
Chromatin Assembly and Disassembly
-
Chromosomal Proteins, Non-Histone / genetics
-
Chromosomal Proteins, Non-Histone / metabolism
-
DNA-Binding Proteins / genetics
-
DNA-Binding Proteins / metabolism
-
Drosophila melanogaster / embryology
-
Drosophila melanogaster / physiology
-
Gene Rearrangement
-
Homeodomain Proteins / genetics
-
Homeodomain Proteins / metabolism*
-
Nuclear Proteins / genetics
-
Nuclear Proteins / metabolism
-
Nucleosomes / metabolism
-
Plasmids / genetics
-
Plasmids / metabolism
-
Recombinant Fusion Proteins / genetics
-
Recombinant Fusion Proteins / metabolism
-
Recombination, Genetic*
-
Trans-Activators / genetics
-
Trans-Activators / metabolism
-
Transcription Factors / genetics
-
Transcription Factors / metabolism
-
Transcription, Genetic*
Substances
-
Chromatin
-
Chromosomal Proteins, Non-Histone
-
DNA-Binding Proteins
-
Gal-VP16
-
Homeodomain Proteins
-
Nuclear Proteins
-
Nucleosomes
-
RAG2 protein, human
-
Recombinant Fusion Proteins
-
SWI-SNF-B chromatin-remodeling complex
-
Trans-Activators
-
Transcription Factors
-
RAG-1 protein