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piRNA-guided slicing specifies transcripts for Zucchini-dependent, phased piRNA biogenesis
Spreading small RNAs to protect the genome
In animals, PIWI-interacting RNAs (piRNAs) are small noncoding RNAs that protect our germ lines from the ravages of transposons. To do this, piRNAs target and cleave transposon RNAs. Synthesis of piRNA is initiated by a cut made in a long, single-stranded precursor RNA. The piRNAs can also undergo a self-perpetuating amplification cycle (see the Perspective by Siomi and Siomi). Han et al. and Mohn et al. now reveal that piRNA biogenesis can also spread in a strictly phased manner from the site of initial piRNA formation. Spreading piRNA synthesis greatly increases their sequence diversity, potentially helping them to target endogenous and novel transposons more effectively.
Abstract
In animal gonads, PIWI-clade Argonaute proteins repress transposons sequence-specifically via bound Piwi-interacting RNAs (piRNAs). These are processed from single-stranded precursor RNAs by largely unknown mechanisms. Here we show that primary piRNA biogenesis is a 3′-directed and phased process that, in the Drosophila germ line, is initiated by secondary piRNA-guided transcript cleavage. Phasing results from consecutive endonucleolytic cleavages catalyzed by Zucchini, implying coupled formation of 3′ and 5′ ends of flanking piRNAs. Unexpectedly, Zucchini also participates in 3′ end formation of secondary piRNAs. Its function can, however, be bypassed by downstream piRNA-guided precursor cleavages coupled to exonucleolytic trimming. Our data uncover an evolutionarily conserved piRNA biogenesis mechanism in which Zucchini plays a central role in defining piRNA 5′ and 3′ ends.
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Science
Volume 348 | Issue 6236
15 May 2015
15 May 2015
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Copyright © 2015, American Association for the Advancement of Science.
Submission history
Received: 20 October 2014
Accepted: 27 February 2015
Published in print: 15 May 2015
Acknowledgments
We thank J. Gokcezade for fly injections, Harvard TRiP and Bloomington stock centers for flies, D. Jurczak for bioinformatics support, the Campus Science Support Facilities Next Generation Sequencing unit for Illumina sequencing, and P. Zamore for sharing unpublished data. This work was supported by the Austrian Academy of Sciences, the European Community (European Research Council grant 260711EU), the Austrian Science Fund (grant Y 510-B12), and Swiss National Science Foundation and Human Frontier Science Program fellowships to F.M. Sequencing data sets are deposited at Gene Expression Omnibus (accession numbers GSE64802 and GSE55842).
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