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Footprint analysis of the RAG protein recombination signal sequence complex for V(D)J type recombination
- Source :
- Molecular and cellular biology. 18(1)
- Publication Year :
- 1998
-
Abstract
- We have studied the interaction between recombination signal sequences (RSSs) and protein products of the truncated forms of recombination-activating genes (RAG) by gel mobility shift, DNase I footprinting, and methylation interference assays. Methylation interference with dimethyl sulfate demonstrated that binding was blocked by methylation in the nonamer at the second-position G residue in the bottom strand and at the sixth- and seventh-position A residues in the top strand. DNase I footprinting experiments demonstrated that RAG1 alone, or even a RAG1 homeodomain peptide, gave footprint patterns very similar to those obtained with the RAG1-RAG2 complex. In the heptamer, partial methylation interference was observed at the sixth-position A residue in the bottom strand. In DNase I footprinting, the heptamer region was weakly protected in the bottom strand by RAG1. The effects of RSS mutations on RAG binding were evaluated by DNA footprinting. Comparison of the RAG-RSS footprint data with the published Hin model confirmed the notion that sequence-specific RSS-RAG interaction takes place primarily between the Hin domain of the RAG1 protein and adjacent major and minor grooves of the nonamer DNA.
- Subjects :
- Genes, RAG-1
DNA Footprinting
Receptors, Antigen, T-Cell
DNA footprinting
Biology
DNA-binding protein
chemistry.chemical_compound
Mice
Recombination signal sequences
Animals
Humans
Molecular Biology
VDJ Recombinases
Homeodomain Proteins
Recombination, Genetic
DNase-I Footprinting
Nuclear Proteins
hemic and immune systems
Cell Biology
Methylation
Molecular biology
DNA Dynamics and Chromosome Structure
DNA-Binding Proteins
chemistry
DNA Nucleotidyltransferases
DNase footprinting assay
DNA
Recombination
Subjects
Details
- ISSN :
- 02707306
- Volume :
- 18
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Molecular and cellular biology
- Accession number :
- edsair.doi.dedup.....290ef09647122ab2ca70e672ad41587e