|
| Status |
Public on May 28, 2015 |
| Title |
Krimper enforces an antisense bias on piRNA pools by binding AGO3 in the Drosophila germline |
| Organism |
Drosophila melanogaster |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
Other
|
| Summary |
Piwi-interacting RNAs (piRNAs) suppress transposon activity in animal germ cells. In the Drosophila ovary, primary Aubergine (Aub)-bound antisense piRNAs initiate the ping-pong cycle to produce secondary AGO3-bound sense piRNAs. This increases the number of secondary Aub-bound antisense piRNAs that can act to destroy transposon mRNAs. Here we show that Krimper (Krimp), a Tudor-domain protein, directly interacts with piRNA-free AGO3 to promote symmetrical dimethylarginine (sDMA) modification, ensuring sense piRNA-loading onto sDMA-modified AGO3. In aub mutant ovaries, AGO3 associates with ping-pong signature piRNAs, suggesting AGO3’s compatibility with primary piRNA loading. Krimp sequesters ectopically expressed AGO3 within Krimp bodies in cultured ovarian somatic cells (OSCs), in which only the primary piRNA pathway operates. Upon krimp-RNAi in OSCs, AGO3 loads with piRNAs, further showing the capacity of AGO3 for primary piRNA loading. We propose that Krimp enforces an antisense bias on piRNA pools by binding AGO3 and blocking its access to primary piRNAs.
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| Overall design |
In order to investigate function of Krimp in piRNA pathway, sequencing of Piwi subfamily protein associated small RNAs was performed using adult Drosophila ovaries and Ovarian Somatic Cells (OSCs) depleted for Krimp or Aub.
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| Contributor(s) |
Sato K, Iwasaki YW, Siomi MC, Siomi H |
| Citation(s) |
26212455 |
| Submission date |
May 27, 2015 |
| Last update date |
Jul 03, 2021 |
| Contact name |
Yuka W. Iwasaki |
| E-mail(s) |
iwasaki@keio.jp
|
| Organization name |
Keio University School of Medicine
|
| Department |
Department of Molecular Biology
|
| Lab |
Siomi Lab
|
| Street address |
35 Shinanomachi, Shinjuku-ku,
|
| City |
Tokyo |
| ZIP/Postal code |
160-8582 |
| Country |
Japan |
| |
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| Platforms (2) |
| GPL13304 |
Illumina HiSeq 2000 (Drosophila melanogaster) |
| GPL16479 |
Illumina MiSeq (Drosophila melanogaster) |
|
| Samples (12)
|
| GSM1697229 |
total small RNA from krimp heterozygous (control) ovaries |
| GSM1697230 |
Piwi associated small RNA from krimp heterozygous (control) ovaries |
| GSM1697231 |
Aub associated small RNA from krimp heterozygous (control) ovaries |
| GSM1697232 |
AGO3 associated small RNA from krimp heterozygous (control) ovaries |
| GSM1697233 |
total small RNA from krimp mutant ovaries |
| GSM1697234 |
Piwi associated small RNA from krimp mutant ovaries |
| GSM1697235 |
Aub associated small RNA from krimp mutant ovaries |
| GSM1697236 |
AGO3 associated small RNA from OR (control) ovaries |
| GSM1697237 |
AGO3 associated small RNA from aub mutant ovaries |
| GSM1697238 |
ectopically expressed myc-Piwi associated small RNA from krimp knockdown OSC |
| GSM1697239 |
ectopically expressed myc-Aub associated small RNA from krimp knockdown OSC |
| GSM1697240 |
ectopically expressed myc-AGO3 associated small RNA from krimp knockdown OSC |
|
| Relations |
| BioProject |
PRJNA285096 |
| SRA |
SRP058770 |
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