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Sensitized piRNA reporter identifies multiple RNA processing factors involved in piRNA-mediated gene silencing

Jordan Brown, View ORCID ProfileDonglei Zhang, Wenjun Chen, View ORCID ProfileHeng-Chi Lee
doi: https://doi.org/10.1101/2023.01.22.525052
Jordan Brown
1Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637, USA
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Donglei Zhang
1Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637, USA
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Wenjun Chen
1Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637, USA
2Department of Laboratory Medicine, Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China
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Heng-Chi Lee
1Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637, USA
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  • ORCID record for Heng-Chi Lee
  • For correspondence: hengchilee@uchicago.edu
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Abstract

Metazoans guard their germlines against transposons and other foreign transcripts with PIWI-interacting RNAs (piRNAs). Due to the robust heritability of the silencing initiated by piRNAs in C.elegans, previous screens using Caenorhabditis elegans were strongly biased to uncover members of this pathway in the maintenance process but not in the initiation process. To identify novel piRNA pathway members, we have utilized a sensitized reporter strain which detects defects in initiation, amplification, or regulation of piRNA silencing. Using our reporter, we have identified Integrator complex subunits, nuclear pore components, protein import components, and pre-mRNA splicing factors as essential for piRNA-mediated gene silencing. We found the snRNA processing cellular machine termed the Integrator complex is required for both type I and type II piRNA production. Notably, we identified a role for nuclear pore and nucleolar components in promoting the perinuclear localization of anti-silencing CSR-1 Argonaute, as well as a role for Importin factor IMA-3 in nuclear localization of silencing Argonaute HRDE-1. Together, we have shown that piRNA silencing is dependent on evolutionarily ancient RNA processing machinery that has been co-opted to function in the piRNA mediated genome surveillance pathway.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted January 22, 2023.
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Sensitized piRNA reporter identifies multiple RNA processing factors involved in piRNA-mediated gene silencing
Jordan Brown, Donglei Zhang, Wenjun Chen, Heng-Chi Lee
bioRxiv 2023.01.22.525052; doi: https://doi.org/10.1101/2023.01.22.525052
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Sensitized piRNA reporter identifies multiple RNA processing factors involved in piRNA-mediated gene silencing
Jordan Brown, Donglei Zhang, Wenjun Chen, Heng-Chi Lee
bioRxiv 2023.01.22.525052; doi: https://doi.org/10.1101/2023.01.22.525052

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