Two Piwi proteins, Xiwi and Xili, are expressed in the Xenopus female germline.
The Argonaute superfamily is a large family of RNA-binding proteins involved in gene regulation mediated by small noncoding RNA and characterized by the presence of PAZ and PIWI domains. The family consists of two branches, the Ago and the Piwi clade. Piwi proteins bind to 21-30-nucleotide-long Piwi-interacting RNAs (piRNAs), which map primarily to transposons and repeated sequence elements. Piwi/piRNAs are important regulators of gametogenesis and have been proposed to play roles in transposon silencing, DNA methylation, transcriptional silencing, and/or post-transcriptional control of translation and RNA stability. Most reports to date have concentrated on the Piwi family members in the male germline. We have identified four Piwi proteins in Xenopus and demonstrate that two, namely, Xiwi1b and Xili, are expressed in the oocyte and early embryo. Xiwi1 and Xili are predominantly found in small, separate complexes, and we do not detect significant interaction of Piwi proteins with the cap-binding complex. Putative nuclear localization and export signals were identified in Xiwi1 and Xili, supporting our observation that Xiwi1, but not Xili, is a nucleo-cytoplasmic protein. Furthermore, by immunoprecipitation of small RNAs, we establish Xiwi1 as a bona fide Piwi protein. These results suggest that the Piwi/piRNA pathway is active in translationally repressed oocytes. This is a significant finding as the Xenopus model provides an excellent tool to study post-transcriptional mechanisms.
Wilczynska
Anna
A
Department of Biochemistry, University of Cambridge, United Kingdom.
Minshall
Nicola
N
Armisen
Javier
J
Miska
Eric A
EA
Standart
Nancy
N
D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D047108:Embryonic Development; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000378:metabolism; D009865:Oocytes / Q000254:growth & development / Q000378:metabolism*; D009866:Oogenesis; D034741:RNA, Small Interfering / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D014981:Xenopus / Q000196:embryology / Q000378:metabolism / Q000502:physiology*; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism*
D046912:Multiprotein Complexes; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D029867:Xenopus Proteins