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Zucchini-dependent piRNA processing is triggered by recruitment to the cytoplasmic processing machinery

Rogers, Alicia K. and Situ, Kathy and Perkins, Edward M. and Fejes Tóth, Katalin (2017) Zucchini-dependent piRNA processing is triggered by recruitment to the cytoplasmic processing machinery. Genes and Development, 31 (18). pp. 1858-1869. ISSN 0890-9369. PMCID PMC5695087. https://resolver.caltech.edu/CaltechAUTHORS:20171016-154844891

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Abstract

The piRNA pathway represses transposable elements in the gonads and thereby plays a vital role in protecting the integrity of germline genomes of animals. Mature piRNAs are processed from longer transcripts, piRNA precursors (pre-piRNAs). In Drosophila, processing of pre-piRNAs is initiated by piRNA-guided Slicer cleavage or the endonuclease Zucchini (Zuc). As Zuc does not have any sequence or structure preferences in vitro, it is not known how piRNA precursors are selected and channeled into the Zuc-dependent processing pathway. We show that a heterologous RNA that lacks complementary piRNAs is processed into piRNAs upon recruitment of several piRNA pathway factors. This processing requires Zuc and the helicase Armitage (Armi). Aubergine (Aub), Argonaute 3 (Ago3), and components of the nuclear RDC complex, which are required for normal piRNA biogenesis in germ cells, are dispensable. Our approach allows discrimination of proteins involved in the transcription and export of piRNA precursors from components required for the cytoplasmic processing steps. piRNA processing correlates with localization of the substrate RNA to nuage, a distinct membraneless cytoplasmic compartment, which surrounds the nucleus of germ cells, suggesting that sequestration of RNA to this subcellular compartment is both necessary and sufficient for selecting piRNA biogenesis substrates.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1101/gad.303214.117DOIArticle
http://genesdev.cshlp.org/content/31/18/1858PublisherArticle
http://genesdev.cshlp.org/content/31/18/1858/suppl/DC1PublisherSupplemental Material
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5695087/PubMed CentralArticle
ORCID:
AuthorORCID
Fejes Tóth, Katalin0000-0001-6558-2636
Additional Information:© 2017 Rogers et al.; Published by Cold Spring Harbor Laboratory Press. This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. Received June 23, 2017; revised version accepted September 13, 2017. Published in Advance October 11, 2017. We are very grateful to Alexei Aravin for discussions and critical reading and editing of the manuscript. We thank members of the Fejes Toth and Aravin laboratoriess for discussions and suggestions, and Maria Ninova for assistance with data analysis. We thank Howard Lipshitz for critical reading and comments on the manuscript. We are grateful to the Brennecke laboratory, the Bloomington Stock Center, and the Kyoto Stock Center for fly stocks. Stocks obtained from the Bloomington Drosophila Stock Center (National Institutes of Health [NIH] P400D018537) and the Drosophila Genomics Resource Center (NIH 2P40OD010949) were used in this study. We are grateful to Igor Antoschechkin at the Millard and Muriel Jacobs Genetics and Genomics Laboratory at California Institute of Technology for sequencing, and the California Institute of Technology Biological Imaging Facility for support with confocal imaging. The Vasa antibody developed by A.C. Spradling and D. Williamswas obtained from the Developmental Studies Hybridoma Bank, created by the National Institute of Child Health and Human Development of the NIH and maintained at The University of Iowa Department of Biology. This work was supported by a grant from the NIH (R01GM110217), and the Ellison Medical Foundation award to K.F.T. A.K.R. is a National Science Foundation (NSF) Graduate Research Fellowship Program (GRFP) fellow. This material is based on work supported by the NSF GRFP (DGE-1144469). Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the National Science Foundation. A.K.R. and K.F.T. designed and executed the experiments. Flies were generated by A.K.R., E.M.P., and K.S. The manuscript was written by K.F.T. and edited by Alexei Aravin, A.K.R., and K.S. Accession numbers: High-throughput sequencing data for small RNA-seq experiments are available through Gene Expression Omnibus (GSE102961).
Funders:
Funding AgencyGrant Number
NIHP400D018537
NIH2P40OD010949
NIHR01GM110217
Ellison Medical FoundationUNSPECIFIED
NSF Graduate Research FellowshipDGE-1144469
Subject Keywords:piRNA biogenesis; nuage; Zucchini; tethering; Piwi; RNA sequestration
Issue or Number:18
PubMed Central ID:PMC5695087
Record Number:CaltechAUTHORS:20171016-154844891
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20171016-154844891
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:82392
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:17 Oct 2017 21:38
Last Modified:09 Mar 2020 13:18

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