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Partha P. Das,1,2,3 Marloes P. Bagijn,1,4 Leonard D. Goldstein,4,5,7 Julie R. Woolford,1,2 Nicolas J. Lehrbach,1,2 Alexandra Sapetschnig,1,2 Heeran R. Buhecha,1,2 Michael J. Gilchrist,1 Kevin L. Howe,5 Rory Stark,5 Nik Matthews,5 Eugene Berezikov,6 René F. Ketting,6 Simon Tavaré,4,5,7 and Eric A. Miska 1,2,
1 Wellcome Trust Cancer Research UK Gurdon Institute, University of Cambridge, The Henry Wellcome Building of Cancer and Developmental Biology, Tennis Court Road, Cambridge CB2 1QN, UK
2 Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1GA, UK
3 PhD Program, Faculty of Biology, GZMB, Georg-August-Universität Göttingen, Justus-von-Liebig-Weg 11, 37077 Göttingen, Germany
4 Department of Oncology, University of Cambridge, Hills Road, Cambridge CB2 2XZ, UK
5 Cancer Research UK, Cambridge Research Institute, Li Ka-Shing Centre, Robinson Way, Cambridge CB2 0RE, UK
6 Hubrecht Institute, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands
7 Department of Applied Mathematics and Theoretical Physics, University of Cambridge, Centre for Mathematical Sciences, Wilberforce Road, Cambridge CB3 0WA, UK
The Piwi proteins of the Argonaute superfamily are required for normal germline development in Drosophila, zebrafish, and mice and associate with 24–30 nucleotide RNAs termed piRNAs. We identify a class of 21 nucleotide RNAs, previously named 21U-RNAs, as the piRNAs of C. elegans. Piwi and piRNA expression is restricted to the male and female germline and independent of many proteins in other small-RNA pathways, including DCR-1. We show that Piwi is specifically required to silence Tc3, but not other Tc/mariner DNA transposons. Tc3 excision rates in the germline are increased at least 100-fold in piwi mutants as compared to wild-type. We find no evidence for a Ping-Pong model for piRNA amplification in C. elegans. Instead, we demonstrate that Piwi acts upstream of an endogenous siRNA pathway in Tc3 silencing. These data might suggest a link between piRNA and siRNA function.
