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| Status |
Public on Oct 22, 2021 |
| Title |
Immunoglobulin enhancers increase RNA polymerase 2 stalling at somatic hypermutation target sequences [GRO-seq] |
| Organism |
Gallus gallus |
| Experiment type |
Other
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| Summary |
Somatic hypermutation (SHM) drives the genetic diversity of immunoglobulin genes in activated B cells and supports the generation of antibodies with increased affinity for antigen. SHM is targeted to Ig genes by their enhancers (DIVACs; diversification activators) but how the enhancers mediate this activity is unknown. We show that DIVACs that strongly stimulate SHM increase the phosphorylation of RNA polymerase 2 (Pol2) and Pol2 occupancy in the mutating gene with little or no accompanying increase in elongation-competent Pol2 or production of full length transcripts, indicating the accumulation of stalled Pol2. DIVAC-induced stalling is weakly associated with an increase in the detection of single-stranded DNA bubbles in the mutating target gene. We did not find evidence for anti-sense transcription or an association of H3K27ac with DIVAC activity in the mutating gene. These findings argue for a connection between Pol2 stalling and cis-acting targeting elements in the context of SHM and thus define a mechanistic basis for locus-specific targeting of SHM in the genome. Our results suggest that DIVACs make the target gene a suitable platform for AID-mediated mutation without the need for increasing transcriptional output.
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| Overall design |
2 samples of GRO-seq profiles in DT40 cell line
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| Contributor(s) |
Tarsalainen A |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| |
| Submission date |
Jul 15, 2021 |
| Last update date |
Oct 23, 2021 |
| Contact name |
Yaakov Maman |
| Organization name |
Bar-Ilan University
|
| Department |
Azrieli Faculty of Medicine
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| Lab |
Genome Instability & Cancer
|
| Street address |
Henrietta Szold 8
|
| City |
Safed |
| ZIP/Postal code |
1311502 |
| Country |
Israel |
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| Platforms (1) |
| GPL19787 |
Illumina NextSeq 500 (Gallus gallus) |
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| Samples (2) |
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| This SubSeries is part of SuperSeries: |
| GSE180178 |
Immunoglobulin enhancers increase RNA polymerase 2 stalling at somatic hypermutation target sequences |
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| Relations |
| BioProject |
PRJNA746967 |
| SRA |
SRP328505 |
