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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Apr 09, 2015 |
| Title |
MIWI and piRNA-mediated cleavage of messenger RNAs in mouse testes |
| Organism |
Mus musculus |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
Expression profiling by high throughput sequencing
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| Summary |
The piRNA machinery is known for its role in mediating epigenetic silencing of transposons. Recent studies suggest that this function also involves piRNA-guided cleavage of transposon-derived transcripts. As many piRNAs also appear to have the capacity to target diverse mRNAs, this raises the intriguing possibility that piRNAs may act extensively as siRNAs to degrade specific mRNAs. To directly test this hypothesis, we compared mouse PIWI (MIWI)-associated piRNAs with experimentally identified cleaved mRNA fragments from mouse testes, and observed cleavage sites that predominantly occur at position 10 from the 5' end of putative targeting piRNAs. We also noted strong biases for U and A residues at nucleotide positions 1 and 10, respectively, in both piRNAs and mRNA fragments, features that resemble the pattern of piRNA amplification by the 'ping-pong' cycle. Through mapping of MIWI-RNA interactions by CLIP-seq and gene expression profiling, we found that many potential piRNA-targeted mRNAs directly interact with MIWI and show elevated expression levels in the testes of Miwi catalytic mutant mice. Reporter-based assays further revealed the importance of base pairing between piRNAs and mRNA targets and the requirement for both the slicer activity and piRNA-loading ability of MIWI in piRNA-mediated target repression. Importantly, we demonstrated that proper turnover of certain key piRNA targets is essential for sperm formation. Together, these findings reveal the siRNA-like function of the piRNA machinery in mouse testes and its central requirement for male germ cell development and maturation.
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| Overall design |
CLIP-Seq (Crosslinking Immunoprecipitation coupled with high-throughput sequencing) experiments targeting Miwi in isolated round spermatids from mouse testis.
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| Contributor(s) |
Zhang P, Kang J, Gou L, Wang J, Xue Y, Skogerboe G, Dai P, Huang D, Chen R, Fu X, Liu M, He S |
| Citation(s) |
25582079, 26677964 |
| Submission date |
Apr 08, 2015 |
| Last update date |
May 15, 2019 |
| Contact name |
Jiao Yuan |
| E-mail(s) |
yuan2006jiao@163.com
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| Organization name |
Institute of Biophysics, Chinese Academy of Sciences
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| Street address |
15 Datun Road, Chaoyang District
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| City |
Beijing |
| ZIP/Postal code |
100101 |
| Country |
China |
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| Platforms (1) |
| GPL9185 |
Illumina Genome Analyzer (Mus musculus) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA280630 |
| SRA |
SRP056988 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE67683_RAW.tar |
87.7 Mb |
(http)(custom) |
TAR (of FA) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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