Journal Article

Miwi catalysis is required for piRNA amplification-independent LINE1 transposon silencing

Chuma, Shinichiro, Hosokawa, Mihoko, Sachidanandam, Ravi, Reuter, Michael, Pillai, Ramesh S., Antony, Claude, Shah, Hardik, Berninger, Philipp, Funaya, Charlotta

2011

Overview

Piwi protein Miwi is shown to be a small RNA-guided RNase in mice; disrupting the catalytic activity of Miwi results in increased accumulation of LINE1 retrotransposon transcripts and male infertility. Transposon silencing by Piwi proteins The combination of Piwi proteins and their associated Piwi-interacting RNAs (piRNAs) mediates epigenetic transposon silencing in animal germlines. Piwi proteins are predicted to be endonucleases, but the significance of this activity had not been demonstrated in vivo . The laboratories of Dónal O'Carroll and Ramesh Pillai have now made mouse models in which residues expected to be critical for nuclease activity in the three mouse Piwi homologues, Mili, Miwi and Miwi2, are mutated. The mutant mice show phenotypic differences. The Mili and Miwi mutants are defective in piRNA production, transposon silencing and fertility, whereas the Miwi2 mutant has normal piRNA levels, seems to undergo piRNA amplification and silences transposons. These studies highlight distinctions between the murine enzymes responsible for piRNA biogenesis. Repetitive-element-derived Piwi-interacting RNAs (piRNAs) 1 , 2 act together with Piwi proteins Mili (also known as Piwil2) and Miwi2 (also known as Piwil4) in a genome defence mechanism that initiates transposon silencing via DNA methylation in the mouse male embryonic germ line. This silencing depends on the participation of the Piwi proteins in a slicer-dependent piRNA amplification pathway and is essential for male fertility 3 , 4 . A third Piwi family member, Miwi (also known as Piwil1), is expressed in specific postnatal germ cells and associates with a unique set of piRNAs of unknown function 5 , 6 , 7 . Here we show that Miwi is a small RNA-guided RNase (slicer) that requires extensive complementarity for target cleavage in vitro . Disruption of its catalytic activity in mice by a single point mutation causes male infertility, and mutant germ cells show increased accumulation of LINE1 retrotransposon transcripts. We provide evidence for Miwi slicer activity directly cleaving transposon messenger RNAs, offering an explanation for the continued maintenance of repeat-derived piRNAs long after transposon silencing is established in germline stem cells. Furthermore, our study supports a slicer-dependent silencing mechanism that functions without piRNA amplification. Thus, Piwi proteins seem to act in a two-pronged mammalian transposon silencing strategy: one promotes transcriptional repression in the embryo, the other reinforces silencing at the post-transcriptional level after birth.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/208/726/2001/1428 / 631/337/176/2016 / 631/337/384/2054 / 631/45/607/1164 / Animals / Argonaute Proteins - deficiency / Argonaute Proteins - genetics / Argonaute Proteins - metabolism / Biocatalysis / Biological and medical sciences / Biosynthesis / Catalysis / DNA methylation / DNA Transposable Elements - genetics / Fertility / Fundamental and applied biological sciences. Psychology / Gene Silencing / Genetic aspects / Genetic research / Genetic transcription / Genic rearrangement. Recombination. Transposable element / Genomes / Humanities and Social Sciences / Infertility / Infertility, Male - genetics / letter / Long Interspersed Nucleotide Elements - genetics / Male / Mice / Molecular and cellular biology / Molecular genetics / multidisciplinary / Mutation - genetics / Physiological aspects / Proteins / RNA / RNA, Messenger - genetics / RNA, Messenger - metabolism / RNA, Small Interfering - biosynthesis / RNA, Small Interfering - genetics / Rodents / Science / Science (multidisciplinary) / Spermatogenesis - genetics / Stem cells / Substrate Specificity / Transposons