title: MIWI and piRNA-mediated cleavage of messenger RNAs in mouse testes.
abstract: 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.
authors.affiliation: Key Laboratory of the Zoological Systematics and Evolution, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.affiliation: 1] Center for RNA Research, State Key Laboratory of Molecular Biology, University of Chinese Academy of Sciences, Shanghai 200031, China [2] Shanghai Key Laboratory of Molecular Andrology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Jun-Yan
authors.initials: JY
authors.lastname: Kang
authors.affiliation: 1] Center for RNA Research, State Key Laboratory of Molecular Biology, University of Chinese Academy of Sciences, Shanghai 200031, China [2] Shanghai Key Laboratory of Molecular Andrology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Lan-Tao
authors.initials: LT
authors.lastname: Gou
authors.affiliation: Key Laboratory of the Zoological Systematics and Evolution, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Jiajia
authors.initials: J
authors.lastname: Wang
authors.affiliation: 1] State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China [2] Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093-0651, USA.
authors.forename: Yuanchao
authors.initials: Y
authors.lastname: Xue
authors.affiliation: Laboratory of Bioinformatics and Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Geir
authors.initials: G
authors.lastname: Skogerboe
authors.affiliation: 1] Center for RNA Research, State Key Laboratory of Molecular Biology, University of Chinese Academy of Sciences, Shanghai 200031, China [2] Shanghai Key Laboratory of Molecular Andrology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Dai
authors.affiliation: Key Laboratory of the Zoological Systematics and Evolution, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Da-Wei
authors.initials: DW
authors.lastname: Huang
authors.affiliation: Laboratory of Bioinformatics and Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Runsheng
authors.initials: R
authors.lastname: Chen
authors.affiliation: 1] State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China [2] Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093-0651, USA.
authors.forename: Xiang-Dong
authors.initials: XD
authors.lastname: Fu
authors.affiliation: 1] Center for RNA Research, State Key Laboratory of Molecular Biology, University of Chinese Academy of Sciences, Shanghai 200031, China [2] Shanghai Key Laboratory of Molecular Andrology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Mo-Fang
authors.initials: MF
authors.lastname: Liu
authors.affiliation: Key Laboratory of the Zoological Systematics and Evolution, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Shunmin
authors.initials: S
authors.lastname: He
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D060565:Argonaute Proteins / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005092:Exonucleases / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D059367:RNA Cleavage; D012333:RNA, Messenger / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D013091:Spermatogenesis; D013094:Spermatozoa / Q000166:cytology; D013737:Testis / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D060565:Argonaute Proteins; D004268:DNA-Binding Proteins; C448522:Piwil1 protein, mouse; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C491866:Xrcc2 protein, mouse; D005092:Exonucleases; C559924:Rad1 protein, mouse