Proximity labeling identifies LOTUS domain proteins that promote the formation of perinuclear germ granules in C. elegans

  1. Ian F Price
  2. Hannah L Hertz
  3. Benjamin Pastore
  4. Jillian Wagner
  5. Wen Tang  Is a corresponding author
  1. Ohio State University, United States

Abstract

The germ line produces gametes that transmit genetic and epigenetic information to the next generation. Maintenance of germ cells and development of gametes require germ granules-well-conserved membraneless and RNA-rich organelles. The composition of germ granules is elusive owing to their dynamic nature and their exclusive expression in the germ line. Using C. elegans germ granule, called P granule, as a model system, we employed a proximity-based labeling method in combination with mass spectrometry to comprehensively define its protein components. This set of experiments identified over 200 proteins, many of which contain intrinsically disordered regions. An RNAi-based screen identified factors that are essential for P granule assembly, notably EGGD-1 and EGGD-2, two putative LOTUS-domain proteins. Loss of eggd-1 and eggd-2 results in separation of P granules from the nuclear envelope, germline atrophy and reduced fertility. We show that intrinsically disordered regions of EGGD-1 are required to anchor EGGD-1 to the nuclear periphery while its LOTUS domains are required to promote perinuclear localization of P granules. Together, our work expands the repertoire of P granule constituents and provides new insights into the role of LOTUS-domain proteins in germ granule organization.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file.Source data files are uploaded to Dryad: https://doi.org/10.5061/dryad.q2bvq83k9Scripts for data analysis are uploaded to Githubhttps://github.com/benpastore/TurboID

The following data sets were generated

Article and author information

Author details

  1. Ian F Price

    Ohio State University, Columbus, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Hannah L Hertz

    Ohio State University, Columbus, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Benjamin Pastore

    Ohio State University, Columbus, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Jillian Wagner

    Ohio State University, Columbus, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Wen Tang

    Ohio State University, Columbus, United States
    For correspondence
    tang.542@osu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6684-5258

Funding

National Institutes of Health (R00GM124460)

  • Wen Tang

National Institutes of Health (R35GM142580)

  • Wen Tang

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Michael Buszczak, University of Texas Southwestern Medical Center, United States

Version history

  1. Received: July 17, 2021
  2. Preprint posted: July 27, 2021 (view preprint)
  3. Accepted: November 1, 2021
  4. Accepted Manuscript published: November 3, 2021 (version 1)
  5. Version of Record published: November 25, 2021 (version 2)

Copyright

© 2021, Price et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Ian F Price
  2. Hannah L Hertz
  3. Benjamin Pastore
  4. Jillian Wagner
  5. Wen Tang
(2021)
Proximity labeling identifies LOTUS domain proteins that promote the formation of perinuclear germ granules in C. elegans
eLife 10:e72276.
https://doi.org/10.7554/eLife.72276

Share this article

https://doi.org/10.7554/eLife.72276

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