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| Status |
Public on Jul 05, 2012 |
| Title |
UNG shapes the specifity of AID-induced somatic hypermutation in non B cells |
| Organism |
Mus musculus |
| Experiment type |
Other
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| Summary |
Secondary diversification of antibodies through somatic hypermutation (SHM) and class switch recombination (CSR) is a critical component of the immune response. Activation-induced deaminase (AID) initiates both processes by deaminating cytosine residues in immunoglobulin genes. The resulting U:G mismatch can be processed by alternative pathways to give rise to a mutation (SHM) or a DNA double-strand break (CSR). Central to this processing is the activity of uracil-N-glycosylase (UNG), an enzyme normally involved in error-free base excision repair. We used next generation sequencing to analyze the contribution of UNG to the resolution of AID-induced lesions. Loss- and gain-of-function experiments showed that UNG activity can promote both error-prone and high fidelity repair of U:G lesions. Unexpectedly, the balance between these alternative outcomes was influenced by the sequence context of the deaminated cytosine, with individual hotspots exhibiting higher susceptibility to UNG-triggered error-free or error-prone resolution. These results reveal UNG as a new molecular layer that shapes the specificity of AID-induced mutations and may provide new insights into the role of AID in cancer development.
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| Overall design |
Next Generation Sequencing analysis of mutations introduced by AID in non B cells. NIH-3T3 cells were co-transduced with mOrangeSTOP and AID-ER–expressing vectors, together with Ugi (UNG inhibitor), UNG, or empty vector as control (n=3). Transduced cells were cultured in the presence of OHT during 11 d. AID-E58Q-ER vector (catalytically inactive form of AID) was used as a negative control in combination with the previously described constructions (n=3).
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| Contributor(s) |
Pérez-Durán P, Belver L, de Yébenes VG, Delgado P, Pisano DG, Ramiro AR |
| Citation(s) |
22665573 |
| |
| Submission date |
Jul 04, 2012 |
| Last update date |
May 15, 2019 |
| Contact name |
Pablo Perez-Duran |
| Organization name |
Centro Nacional de Investigaciones Cardiovasculares (CNIC)
|
| Street address |
Melchor Fernandez Almagro, 3
|
| City |
Madrid |
| ZIP/Postal code |
28002 |
| Country |
Spain |
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| Platforms (1) |
| GPL11002 |
Illumina Genome Analyzer IIx (Mus musculus) |
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| Samples (18)
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| This SubSeries is part of SuperSeries: |
| GSE39115 |
UNG shapes the specificity of AID-induced somatic hypermutation |
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| Relations |
| BioProject |
PRJNA170048 |
| SRA |
SRP014022 |
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