Novel LOTUS-domain proteins are organizational hubs that recruit C. elegans Vasa to germ granules
- New York University, United States
- McGill University, Canada
- Max Delbrück Center for Molecular Medicine, Germany
- New York University Abu Dhabi, United Arab Emirates
Abstract
We describe MIP-1 and MIP-2, novel paralogous C. elegans germ granule components that interact with the intrinsically disordered MEG-3 protein. These proteins promote P granule condensation, form granules independently of MEG-3 in the postembryonic germ line, and balance each other in regulating P granule growth and localization. MIP-1 and MIP-2 each contain two LOTUS domains and intrinsically disordered regions and form homo- and heterodimers. They bind and anchor the Vasa homolog GLH-1 within P granules and are jointly required for coalescence of MEG-3, GLH-1, and PGL proteins. Animals lacking MIP-1 and MIP-2 show temperature-sensitive embryonic lethality, sterility, and mortal germ lines. Germline phenotypes include defects in stem cell self-renewal, meiotic progression, and gamete differentiation. We propose that these proteins serve as scaffolds and organizing centers for ribonucleoprotein networks within P granules that help recruit and balance essential RNA processing machinery to regulate key developmental transitions in the germ line.
Data availability
All mass spectrometry raw data have been deposited to the PRIDE repository with the dataset identifier PXD012852. All other data generated or analyzed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 2A-C; Figure 2-figure supplement 2; Figure 6A,B; Figure 8E; Figure 9B; Figure 9-figure supplement 1; Figure 10C.
Article and author information
Author details
Kristin C Gunsalus
Funding
New York University Abu Dhabi (ADPHG CGSB)
- Patricia Giselle Cipriani
- Hala Fahs
- Fabio Piano
- Kristin C Gunsalus
New York University Abu Dhabi
- Patricia Giselle Cipriani
- Olivia Bay
- John Zinno
- Michelle Gutwein
- Hin Hark Gan
- George Chung
- Fabio Piano
- Kristin C Gunsalus
Canadian Institutes of Health Research (MOP 123352)
- Vinay K Mayya
- Thomas F Duchaine
Charlotte and Leo Karassik Foundation
- Vinay K Mayya
Bundesministerium für Bildung und Forschung (0315362)
- Jia-Xuan Chen
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Michael Buszczak, University of Texas Southwestern Medical Center, United States
Version history
- Received: July 8, 2020
- Accepted: June 27, 2021
- Accepted Manuscript published: July 5, 2021 (version 1)
- Version of Record published: August 3, 2021 (version 2)
Copyright
© 2021, Cipriani 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.
Metrics
-
- 1,997
- Page views
-
- 191
- Downloads
-
- 7
- Citations
Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Novel LOTUS-domain proteins are organizational hubs that recruit C. elegans Vasa to germ granules
eLife 10:e60833.
https://doi.org/10.7554/eLife.60833
Further reading
-
- Cell Biology
Epidemiological studies have demonstrated that fine particulate matter (PM2.5) is associated with adverse obstetric and postnatal metabolic health outcomes, but the mechanism remains unclear. This study aimed to investigate the toxicological pathways by which PM2.5 damaged placental trophoblasts in vivo and in vitro. We confirmed that PM2.5 induced adverse gestational outcomes such as increased fetal mortality rates, decreased fetal number and weight, damaged placental structure, and increased apoptosis of trophoblasts. Additionally, PM2.5 induced dysfunction of the trophoblast cell line HTR8/SVneo, including in its proliferation, apoptosis, invasion, migration and angiogenesis. Moreover, we comprehensively analyzed the transcriptional landscape of HTR8/SVneo cells exposed to PM2.5 through RNA-Seq and observed that PM2.5 triggered overexpression of pathways involved in oxidative stress and mitochondrial apoptosis to damage HTR8/SVneo cell biological functions through CYP1A1. Mechanistically, PM2.5 stimulated KLF9, a transcription factor identified as binding to CYP1A1 promoter region, which further modulated the CYP1A1-driven downstream phenotypes. Together, this study demonstrated that the KLF9/CYP1A1 axis played a crucial role in the toxic progression of PM2.5 induced adverse pregnancy outcomes, suggesting adverse effects of environmental pollution on pregnant females and putative targeted therapeutic strategies.
-
- Cell Biology
- Microbiology and Infectious Disease
Reverse genetics is key to understanding protein function, but the mechanistic connection between a gene of interest and the observed phenotype is not always clear. Here we describe the use of proximity labeling using TurboID and site-specific quantification of biotinylated peptides to measure changes to the local protein environment of selected targets upon perturbation. We apply this technique, which we call PerTurboID, to understand how the P. falciparum exported kinase, FIKK4.1, regulates the function of the major virulence factor of the malaria causing parasite, PfEMP1. We generated independent TurboID fusions of 2 proteins that are predicted substrates of FIKK4.1 in a FIKK4.1 conditional KO parasite line. Comparing the abundance of site-specific biotinylated peptides between wildtype and kinase deletion lines reveals the differential accessibility of proteins to biotinylation, indicating changes to localization, protein-protein interactions, or protein structure which are mediated by FIKK4.1 activity. We further show that FIKK4.1 is likely the only FIKK kinase that controls surface levels of PfEMP1, but not other surface antigens, on the infected red blood cell under standard culture conditions. We believe PerTurboID is broadly applicable to study the impact of genetic or environmental perturbation on a selected cellular niche.
