|
| Status |
Public on Jun 05, 2015 |
| Title |
Somatic Primary piRNA Biogenesis Driven by Cis-Acting RNA Elements and Trans- Acting Yb |
| Organism |
Drosophila melanogaster |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
|
| Summary |
Primary piRNAs in Drosophila ovarian somatic cells arise from piRNA cluster transcripts and the 3′ UTRs of a subset of mRNAs, including Traffic jam (Tj) mRNA. However, it is unclear how these RNAs are determined as primary piRNA sources. Here, we identify a cis-acting 100-nt fragment in the Tj 3′ UTR that is sufficient for producing artificial piRNAs from unintegrated DNA. These artificial piRNAs were effective in endogenous gene transcriptional silencing. Yb, a core component of primary piRNA biogenesis center Yb bodies, directly bound the Tj-cis-element. Disruption of this interaction markedly reduced piRNA production. Thus, Yb is the trans-acting partner of the Tj-cis-element. Yb-CLIP revealed that Yb-binding correlated with somatic piRNA production but Tj-cis-element downstream sequences produced few artificial piRNAs. Thus, Yb determines primary piRNA sources by two modes of action; primary binding to cis-elements to specify substrates, and secondary binding to downstream regions to increase diversity in piRNA populations.
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| |
|
| Overall design |
HITS-CLIP of Yb in OSCs (Ovarian Somatic Cells) depleted for tj cis-element, and small RNA sequencing of Piwi-piRNAs in OSCs depleted for tj cis-element.
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| |
|
| Contributor(s) |
Ishizu H, Siomi MC |
| Citation(s) |
26166564 |
| Submission date |
Jun 05, 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 |
| |
|
| Platforms (2) |
| GPL13304 |
Illumina HiSeq 2000 (Drosophila melanogaster) |
| GPL16479 |
Illumina MiSeq (Drosophila melanogaster) |
|
| Samples (8)
|
| GSM1704899 |
YbCLIP in tj cis-element deleted OSCs |
| GSM1704900 |
Piwi-associated smRNA from non-deleted OSCs |
| GSM1704901 |
Piwi-associated smRNA from tj cis-element deleted OSCs 1 |
| GSM1704902 |
Piwi-associated smRNA from tj cis-element deleted OSCs 2 |
| GSM1704903 |
Piwi-associated smRNA from EGFP artificial piRNA expressed OSCs |
|
| Relations |
| BioProject |
PRJNA286011 |
| SRA |
SRP059207 |
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