Identification of MIWI-associated Poly(A) RNAs by immunoprecipitation with an anti-MIWI monoclonal antibody

Nishibu, Takahiro; Hayashida, Yukinobu; Tani, Satoshi; Kurono, Sadamu; Kojima‐Kita, Kanako; Ukekawa, Ryo; Kurokawa, Tsutomu; Kuramochi‐Miyagawa, Satomi; Nakano, Toru; Inoue, Kimiko; Honda, Susumu

doi:10.5582/bst.2012.v6.5.248

Cited by 19 publications

(11 citation statements)

References 50 publications

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“…Our finding appears to contradict with recent studies reporting that MIWI binds and translationally represses spermiogenic mRNAs in a piRNA-independent manner in mouse testes 28,47 . Such “piRNA-independent” function might be responsible for protecting target mRNAs in SC and early RS until the next developmental phase.…”

Section: Discussioncontrasting

confidence: 99%

Pachytene piRNAs instruct massive mRNA elimination during late spermiogenesis

et al. 2014

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Spermatogenesis in mammals is characterized by two waves of piRNA expression: one corresponds to classic piRNAs responsible for silencing retrotransponsons and the second wave is predominantly derived from nontransposon intergenic regions in pachytene spermatocytes, but the function of these pachytene piRNAs is largely unknown. Here, we report the involvement of pachytene piRNAs in instructing massive mRNA elimination in mouse elongating spermatids (ES). We demonstrate that a piRNA-induced silencing complex (pi-RISC) containing murine PIWI (MIWI) and deadenylase CAF1 is selectively assembled in ES, which is responsible for inducing mRNA deadenylation and decay via a mechanism that resembles the action of miRNAs in somatic cells. Such a highly orchestrated program appears to take full advantage of the enormous repertoire of diversified targeting capacity of pachytene piRNAs derived from nontransposon intergenic regions. These findings suggest that pachytene piRNAs are responsible for inactivating vast cellular programs in preparation for sperm production from ES.

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Section: Discussioncontrasting

confidence: 99%

Pachytene piRNAs instruct massive mRNA elimination during late spermiogenesis

et al. 2014

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“…Intriguingly, MIWI was shown to associate with the translational machinery as well as with polysomes during early spermiogenesis, leading to speculation that it promotes translation (Lau, 2010). Recent work also suggests that MIWI promotes the stabilization of spermiogenic mRNAs (Nishibu et al, 2012; Vourekas et al, 2012). Interestingly, a protective role for small RNAs and PIWI Argonaute proteins was recently identified in the ciliated protozoan Oxytricha, where piRNAs prevent DNA elimination during genome rearrangement (Fang et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

Argonautes Promote Male Fertility and Provide a Paternal Memory of Germline Gene Expression in C. elegans

Conine

Moresco

et al. 2013

Cell

167

268

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SUMMARY During each life cycle germ cells preserve and pass on both genetic and epigenetic information. In C. elegans, the ALG-3/4 Argonaute proteins are expressed during male gametogenesis and promote male fertility. Here we show that the CSR-1 Argonaute functions with ALG-3/4 to positively regulate target genes required for spermiogenesis. Our findings suggest that ALG-3/4 functions during spermatogenesis to amplify a small-RNA signal that represents an epigenetic memory of male-specific gene expression. CSR-1, which is abundant in mature sperm, appears to transmit this memory to offspring. Surprisingly, in addition to small RNAs targeting male-specific genes, we show that males also harbor an extensive repertoire of CSR-1 small RNAs targeting oogenesis-specific mRNAs. Together these findings suggest that C. elegans sperm transmit not only the genome but also epigenetic binary signals in the form of Argonaute/small-RNA complexes that constitute a memory of gene expression in preceding generations.

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“…Following consideration of the observation that loading of pachytene piRNAs directs MIWI toward the ubiquitin-proteosome pathway for degradation (Zhao et al 2013), an alternative model emerged, which suggested that in the absence of meiotic piRNAs, MIWI was stabilized so it can inhibit translation. In addition to repressing or activating translation, MIWI has also been implicated in promoting RNA stability (Nishibu et al 2012;Vourekas et al 2012). Given that MIWI-associated mRNAs, identified by cross-linking immunoprecipitation (CLIP), were less abundant in Miwi mutants, it was argued that MIWI binding stabilizes bound mRNAs.…”

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confidence: 99%

piRNA-directed cleavage of meiotic transcripts regulates spermatogenesis

et al. 2015

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MIWI catalytic activity is required for spermatogenesis, indicating that piRNA-guided cleavage is critical for germ cell development. To identify meiotic piRNA targets, we augmented the mouse piRNA repertoire by introducing a human meiotic piRNA cluster. This triggered a spermatogenesis defect by inappropriately targeting the piRNA machinery to mouse mRNAs essential for germ cell development. Analysis of such de novo targets revealed a signature for pachytene piRNA target recognition. This enabled identification of both transposable elements and meiotically expressed protein-coding genes as targets of native piRNAs. Cleavage of genic targets began at the pachytene stage and resulted in progressive repression through meiosis, driven at least in part via the ping-pong cycle. Our data support the idea that meiotic piRNA populations must be strongly selected to enable successful spermatogenesis, both driving the response away from essential genes and directing the pathway toward mRNA targets that are regulated by small RNAs in meiotic cells.

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Identification of MIWI-associated Poly(A) RNAs by immunoprecipitation with an anti-MIWI monoclonal antibody

Cited by 19 publications

References 50 publications

Pachytene piRNAs instruct massive mRNA elimination during late spermiogenesis

Pachytene piRNAs instruct massive mRNA elimination during late spermiogenesis

Argonautes Promote Male Fertility and Provide a Paternal Memory of Germline Gene Expression in C. elegans

piRNA-directed cleavage of meiotic transcripts regulates spermatogenesis

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