title: Modulation of host ROS metabolism is essential for viral infection of a bloom-forming coccolithophore in the ocean.
abstract: The cosmopolitan coccolithophore Emiliania huxleyi is a unicellular eukaryotic alga responsible for vast blooms in the ocean. These blooms have immense impact on large biogeochemical cycles and are terminated by a specific large double-stranded DNA E. huxleyi virus (EhV, Phycodnaviridae). EhV infection is accompanied by induction of hallmarks of programmed cell death and production of reactive oxygen species (ROS). Here we characterized alterations in ROS metabolism and explored its role during infection. Transcriptomic analysis of ROS-related genes predicted an increase in glutathione (GSH) and H2O2 production during infection. In accordance, using biochemical assays and specific fluorescent probes we demonstrated the overproduction of GSH during lytic infection. We also showed that H2O2 production, rather than superoxide, is the predominant ROS during the onset of the lytic phase of infection. Using flow cytometry, confocal microscopy and multispectral imaging flow cytometry, we showed that the profound co-production of H2O2 and GSH occurred in the same subpopulation of cells but at different subcellular localization. Positively stained cells for GSH and H2O2 were highly infected compared with negatively stained cells. Inhibition of ROS production by application of a peroxidase inhibitor or an H2O2 scavenger inhibited host cell death and reduced viral production. We conclude that viral infection induced remodeling of the host antioxidant network that is essential for a successful viral replication cycle. This study provides insight into viral replication strategy and suggests the use of specific cellular markers to identify and quantify the extent of active viral infection during E. huxleyi blooms in the ocean.
authors.affiliation: Department of Plant and Environmental Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Uri
authors.initials: U
authors.lastname: Sheyn
authors.affiliation: Department of Plant and Environmental Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Shilo
authors.initials: S
authors.lastname: Rosenwasser
authors.affiliation: Department of Biological Services, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Shifra
authors.identifier: 0000-0001-9604-1939
authors.initials: S
authors.lastname: Ben-Dor
authors.affiliation: Department of Biological Services, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Ziv
authors.initials: Z
authors.lastname: Porat
authors.affiliation: Department of Plant and Environmental Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Assaf
authors.initials: A
authors.lastname: Vardi
mesh_terms: D000975:Antioxidants / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D058087:Haptophyta / Q000502:physiology / Q000821:virology*; D054884:Host-Pathogen Interactions; D006861:Hydrogen Peroxide / Q000378:metabolism; D009792:Oceans and Seas; D010084:Oxidation-Reduction; D017862:Phycodnaviridae / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D000975:Antioxidants; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D005978:Glutathione