Viral activation and recruitment of metacaspases in the unicellular coccolithophore, Emiliania huxleyi.
Lytic viral infection and programmed cell death (PCD) are thought to represent two distinct death mechanisms in phytoplankton, unicellular photoautotrophs that drift with ocean currents. Here, we demonstrate an interaction between autocatalytic PCD and lytic viral infection in the cosmopolitan coccolithophorid, Emiliania huxleyi. Successful infection of E. huxleyi strain 374 with a lytic virus, EhV1, resulted in rapid internal degradation of cellular components, a dramatic reduction in the photosynthetic efficiency (F(v)/F(m)), and an up-regulation of metacaspase protein expression, concomitant with induction of caspase-like activity. Caspase activation was confirmed through in vitro cleavage in cell extracts of the fluorogenic peptide substrate, IETD-AFC, and direct, in vivo staining of cells with the fluorescently labeled irreversible caspase inhibitor, FITC-VAD-FMK. Direct addition of z-VAD-FMK to infected cultures abolished cellular caspase activity and protein expression and severely impaired viral production. The absence of metacaspase protein expression in resistant E. huxleyi strain 373 during EhV1 infection further demonstrated the critical role of these proteases in facilitating viral lysis. Together with the presence of caspase cleavage recognition sequences within virally encoded proteins, we provide experimental evidence that coccolithoviruses induce and actively recruit host metacaspases as part of their replication strategy. These findings reveal a critical role for metacaspases in the turnover of phytoplankton biomass upon infection with viruses and point to coevolution of host-virus interactions in the activation and maintenance of these enzymes in planktonic, unicellular protists.
Bidle
Kay D
KD
Environmental Biophysics and Molecular Ecology Group, Institute of Marine and Coastal Sciences, Rutgers, The State University of New Jersey, 71 Dudley Road, New Brunswick, NJ 08901, USA. bidle@marine.rutgers.edu
Haramaty
Liti
L
Barcelos E Ramos
Joana
J
Falkowski
Paul
P
D017209:Apoptosis; D020169:Caspases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006566:Herpesviridae Infections; D004861:Herpesvirus 1, Equid / Q000472:pathogenicity* / Q000502:physiology / Q000648:ultrastructure; D007700:Kinetics; D010839:Phytoplankton / Q000201:enzymology* / Q000254:growth & development / Q000648:ultrastructure / Q000821:virology*; D012623:Seawater / Q000382:microbiology; D013379:Substrate Specificity; D015854:Up-Regulation; D014775:Virus Activation
D005456:Fluorescent Dyes; D020169:Caspases; D016650:Fluorescein-5-isothiocyanate