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| Status |
Public on Apr 20, 2010 |
| Title |
The In Vivo Pattern of Binding of RAG1 and RAG2 to Antigen Receptor Loci |
| Organism |
Mus musculus |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
The critical initial step in V(D)J recombination, binding of RAG1 and RAG2 to recombination signal sequences flanking antigen receptor V, D, and J gene segments, has not previously been characterized in vivo. Here we demonstrate that RAG protein binding occurs in a highly focal manner to a small region of active chromatin encompassing Igκ and Tcrα J gene segments and Igh and Tcrβ J and J-proximal D gene segments. Formation of these small RAG-bound regions, which we refer to as recombination centers, occurs in a developmental stage- and lineage-specific manner. Each RAG protein is independently capable of specific binding within recombination centers. While RAG1 binding is restricted to regions containing recombination signal sequences, RAG2 binds extremely broadly in a pattern that mirrors that of trimethylated lysine 4 of histone 3. We propose that recombination centers coordinate V(D)J recombination by providing discrete sites within which gene segments are captured for recombination.
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| Overall design |
RAG2 binding was analyzed in wild type, RAG2-/-β, and D708A-RAG1-/-β thymocytes. Histone modification H3K4me3 was analyzed in D708A-RAG1-/-β and wild type thymocytes.
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| Contributor(s) |
Ji Y, Resch W, Corbett E, Yamane A, Casellas R, Schatz DG |
| Citation(s) |
20398922 |
| |
| Submission date |
Apr 05, 2010 |
| Last update date |
May 15, 2019 |
| Contact name |
Seolkyoung Jung |
| Organization name |
NIH
|
| Department |
NIAMS
|
| Lab |
biodata mining and discovery section
|
| Street address |
10 Center Dr
|
| City |
bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
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| Platforms (1) |
| GPL9250 |
Illumina Genome Analyzer II (Mus musculus) |
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| Samples (5)
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| Relations |
| SRA |
SRP002241 |
| BioProject |
PRJNA126295 |
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