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| Status |
Public on Dec 31, 2014 |
| Title |
High throughput sequencing of Siwi and BmAGO3 associated piRNAs from Bombyx mori BmN4 cells |
| Organism |
Bombyx mori |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
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| Summary |
In germ cells, piRNAs are amplified through the Ping-Pong cycle that depends on reciprocal Slicer-mediated target RNA cleavage by two PIWI members. A germ-specific DEAD-box protein Vasa is required for the process. However, Vasa’s function is poorly understood. Here, we show that target RNAs cleaved by a Bombyx mori (silkworm) PIWI, Siwi, remain to be bound with the protein upon cleavage, but are released in the presence of Vasa in B. mori (BmVasa) and ATP. Under normal conditions, BmVasa co-purifies with Siwi, but not with second B. mori PIWI BmAgo3. However, when BmVasa loses the ATP-binding and RNA-unwinding activities, BmVasa avidly associates with Siwi and BmAgo3, which contains transposon transcripts predominantly in sense orientation, the sources of BmAgo3-piRNAs. Without BmVasa, BmAgo3 is devoid of piRNAs. Thus, BmVasa actively releases target RNAs from Siwi, upon its cleavage, to urge BmAgo3-piRNA complex formation in the Ping-Pong cycle, enabling continuous supply of piRNAs in germ cells.
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| Overall design |
small RNA libraries were prepared from Siwi, BmAGO3, and Flag-Siwi immuno-precipitate samples of BmN4 cells
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| Contributor(s) |
Nishida KM, Siomi H, Siomi MC |
| Citation(s) |
25558067 |
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| Submission date |
Jun 04, 2014 |
| Last update date |
Jul 03, 2021 |
| Contact name |
Yuka W. Iwasaki |
| E-mail(s) |
iwasaki@keio.jp
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| Organization name |
Keio University School of Medicine
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| Department |
Department of Molecular Biology
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| Lab |
Siomi Lab
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| Street address |
35 Shinanomachi, Shinjuku-ku,
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| City |
Tokyo |
| ZIP/Postal code |
160-8582 |
| Country |
Japan |
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| Platforms (1) |
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| Samples (5)
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| GSM1533654 |
Flag-Siwi bound piRNAs from Vasa siRNA treated BmN4 |
| GSM1533655 |
Flag-Siwi bound piRNAs from BmAGO3 siRNA treated BmN4 |
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| Relations |
| BioProject |
PRJNA251635 |
| SRA |
SRP042945 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE58221_RAW.tar |
1.2 Mb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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