|
| Status |
Public on Jul 01, 2014 |
| Title |
The MID-PIWI module of Piwi proteins specifies nucleotide- and strand-biases of piRNAs |
| Organism |
Bombyx mori |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
|
| Summary |
Piwi-interacting RNAs (piRNAs) guide Piwi Argonautes to suppress transposon activity in animal gonads. Known piRNA populations are extremely complex, with millions of individual sequences present in a single organism. Despite this complexity, specific Piwi proteins incorporate piRNAs with distinct nucleotide- and transposon strand-biases (antisense or sense) of unknown origin. Here we examined the contribution of structural domains in Piwi proteins towards defining these biases. We report the first crystal structure of the MID domain from a Piwi Argonaute and use docking experiments to show its ability to specify recognition of 5′ uridine (1U-bias) of piRNAs. Mutational analyses reveal the importance of 5’ end-recognition within the MID domain for piRNA biogenesis in vivo. Finally, domain-swapping experiments uncover an unexpected role for the MID-PIWI module of a Piwi protein in dictating the strand-orientation of its bound piRNAs. Our work identifies structural features that allow distinguishing individual Piwi members during piRNA biogenesis
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| Overall design |
Immunoprecipitated small RNA were purified from Bmn4 cells for preparation of high-throughput sequencing libraries.
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| Contributor(s) |
Cora E, Pandey RR, Xiol J, Taylor J, Sachidanandam R, McCarthy AA, Pillai RS |
| Citation(s) |
24757166 |
| |
| Submission date |
Feb 28, 2014 |
| Last update date |
May 15, 2019 |
| Contact name |
Ramesh Pillai |
| E-mail(s) |
ramesh.pillai@unige.ch
|
| Organization name |
University of Geneva
|
| Department |
Department of Molecular Biology
|
| Street address |
30, Quai Ernest-Ansermet
|
| City |
Gneveva |
| ZIP/Postal code |
CH-1211 |
| Country |
Switzerland |
| |
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| Platforms (1) |
| GPL16216 |
Illumina HiSeq 2000 (Bombyx mori) |
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| Samples (7)
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| Relations |
| BioProject |
PRJNA239665 |
| SRA |
SRP039056 |
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