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Toward Structure Analysis of a Huge Algal Virus

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HOMOLA Miroslav BÜTTNER Carina Renate FÜZIK Tibor PLEVKA Pavel

Rok publikování 2020
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Středoevropský technologický institut

Citace
Popis The Emiliania huxleyi virus 86 (EhV-86) is an important agent in terminating regular population explosions of Emiliania huxleyi – the most abundant alga in marine ecosystems. We are attempting to solve the structure of EhV-86 using several cryo-electron microscopy techniques. The unicellular alga Emiliania huxleyi is widely distributed photosynthesizing marine eukaryote, creating regular population explosions called blooms. These algal blooms, visible from space, influence the global climate by absorption of carbon dioxide as E. huxleyi uses this molecule for produc-tion of calcareous plates, covering its surface. The algal bloom collapses usually one week after infection by coccolithoviruses, in most cases by EhV-86. Among the other large double-stranded DNA viruses that infect green algae, EhV-86 differs in possessing a lipid membrane outside of the capsid shell, which it acquires while exiting the algal cell via budding. The membrane and large dimensions of the EhV-86 virus particle make it quite challenging for purification and subsequent 3-dimensional reconstruction. To determine the near-atomic resolution structure of the EhV-86 virion, we use cryo-electron microscopy (cryo-EM). The observed heterogeneity of viral particle morphology pushes reconstruction by so-called single particle analysis, i.e. averaging over many particles – toward tomography, i.e. using only few particles, each imaged at many angles. We also investigate the rep-lication machinery of EhV-86 inside the algal host cell using lamellas – thin cell sections produced by focused ion beam milling. The combination of our results obtained from various cryo-EM approaches will provide a compre-hensive insight into the EhV-86 life cycle and interactions with the Emiliania huxleyi host cell.
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