title: Small GTPases promote actin coat formation on microsporidian pathogens traversing the apical membrane of Caenorhabditis elegans intestinal cells.
abstract: Many intracellular pathogens co-opt actin in host cells, but little is known about these interactions in vivo. We study the in vivo trafficking and exit of the microsporidian Nematocida parisii, which is an intracellular pathogen that infects intestinal cells of the nematode Caenorhabditis elegans. We recently demonstrated that N. parisii uses directional exocytosis to escape out of intestinal cells into the intestinal tract. Here, we show that an intestinal-specific isoform of C. elegans actin called ACT-5 forms coats around membrane compartments that contain single exocytosing spores, and that these coats appear to form after fusion with the apical membrane. We performed a genetic screen for host factors required for actin coat formation and identified small GTPases important for this process. Through analysis of animals defective in these factors, we found that actin coats are not required for pathogen exit although they may boost exocytic output. Later during infection, we find that ACT-5 also forms coats around membrane-bound vesicles that contain multiple spores. These vesicles are likely formed by clathrin-dependent compensatory endocytosis to retrieve membrane material that has been trafficked to the apical membrane as part of the exocytosis process. These findings provide insight into microsporidia interaction with host cells, and provide novel in vivo examples of the manner in which intracellular pathogens co-opt host actin during their life cycle.
authors.affiliation: Division of Biological Sciences, Section of Cell and Developmental Biology, University of California, La Jolla, CA, USA.
authors.forename: Suzannah C
authors.initials: SC
authors.lastname: Szumowski
authors.affiliation: Division of Biological Sciences, Section of Cell and Developmental Biology, University of California, La Jolla, CA, USA.
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Estes
authors.affiliation: Division of Biological Sciences, Section of Cell and Developmental Biology, University of California, La Jolla, CA, USA.
authors.forename: John J
authors.initials: JJ
authors.lastname: Popovich
authors.affiliation: Division of Biological Sciences, Section of Cell and Developmental Biology, University of California, La Jolla, CA, USA.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Botts
authors.affiliation: Division of Biological Sciences, Section of Cell and Developmental Biology, University of California, La Jolla, CA, USA.
authors.forename: Grace
authors.initials: G
authors.lastname: Sek
authors.affiliation: Division of Biological Sciences, Section of Cell and Developmental Biology, University of California, La Jolla, CA, USA.
authors.forename: Emily R
authors.initials: ER
authors.lastname: Troemel
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D017173:Caenorhabditis elegans / Q000382:microbiology*; D004847:Epithelial Cells / Q000382:microbiology*; D005089:Exocytosis; D054884:Host-Pathogen Interactions; D016814:Microsporidia / Q000502:physiology*; D020559:Monomeric GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D000199:Actins; D020559:Monomeric GTP-Binding Proteins