PIWIL3 Forms a Complex with TDRKH in Mammalian Oocytes
Minjie Tan,
Helena T.A. van Tol,
David Rosenkranz,
Elke F. Roovers,
Mirjam J. Damen,
Tom A.E. Stout,
Wei Wu,
Bernard A.J. Roelen
Affiliations
Minjie Tan
Farm Animal Health, Department of Population Health Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM Utrecht, The Netherlands
Helena T.A. van Tol
Farm Animal Health, Department of Population Health Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM Utrecht, The Netherlands
David Rosenkranz
Johannes Gutenberg-University Mainz, Institute of Organismic and Molecular Evolution, Anselm-Franz-von-Bentzel-Weg 7, 55128 Mainz, Germany
Elke F. Roovers
Biology of Non-coding RNA Group, Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz, Germany
Mirjam J. Damen
Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands
Tom A.E. Stout
Equine Sciences, Department Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 112, 3584 CM Utrecht, The Netherlands
Wei Wu
Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands
Bernard A.J. Roelen
Embryology, Anatomy and Physiology, Department Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands
P-element induced wimpy testis (PIWIs) are crucial guardians of genome integrity, particularly in germ cells. While mammalian PIWIs have been primarily studied in mouse and rat, a homologue for the human PIWIL3 gene is absent in the Muridae family, and hence the unique function of PIWIL3 in germ cells cannot be effectively modeled by mouse knockouts. Herein, we investigated the expression, distribution, and interaction of PIWIL3 in bovine oocytes. We localized PIWIL3 to mitochondria, and demonstrated that PIWIL3 expression is stringently controlled both spatially and temporally before and after fertilization. Moreover, we identified PIWIL3 in a mitochondrial-recruited three-membered complex with Tudor and KH domain-containing protein (TDRKH) and poly(A)-specific ribonuclease-like domain containing 1 (PNLDC1), and demonstrated by mutagenesis that PIWIL3 N-terminal arginines are required for complex assembly. Finally, we sequenced the piRNAs bound to PIWIL3-TDRKH-PNLDC1 and report here that about 50% of these piRNAs map to transposable elements, recapitulating the important role of PIWIL3 in maintaining genome integrity in mammalian oocytes.
