Halorubrum pleomorphic virus-6 Membrane Fusion Is Triggered by an S-Layer Component of Its Haloarchaeal Host

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Bignon, E.A.; Chou, K.R.; Roine, E.; Tischler, N.D. Halorubrum pleomorphic virus-6 Membrane Fusion Is Triggered by an S-Layer Component of Its Haloarchaeal Host. Viruses 2022, 14, 254.

Title: Halorubrum pleomorphic virus-6 Membrane Fusion Is Triggered by an S-Layer Component of Its Haloarchaeal Host
Author: Bignon, Eduardo A.; Chou, Kevin R.; Roine, Elina; Tischler, Nicole D.
Publisher: Multidisciplinary Digital Publishing Institute
Date: 2022-01-27
URI: http://hdl.handle.net/10138/340156
Abstract: (1) Background: Haloarchaea comprise extremely halophilic organisms of the Archaea domain. They are single-cell organisms with distinctive membrane lipids and a protein-based cell wall or surface layer (S-layer) formed by a glycoprotein array. Pleolipoviruses, which infect haloarchaeal cells, have an envelope analogous to eukaryotic enveloped viruses. One such member, <i>Halorubrum pleomorphic virus 6</i> (HRPV-6), has been shown to enter host cells through virus-cell membrane fusion. The HRPV-6 fusion activity was attributed to its VP4-like spike protein, but the physiological trigger required to induce membrane fusion remains yet unknown. (2) Methods: We used SDS-PAGE mass spectroscopy to characterize the S-layer extract, established a proteoliposome system, and used R18-fluorescence dequenching to measure membrane fusion. (3) Results: We show that the S-layer extraction by Mg<sup>2+</sup> chelating from the HRPV-6 host, <i>Halorubrum</i> sp. SS7-4, abrogates HRPV-6 membrane fusion. When we in turn reconstituted the S-layer extract from <i>Hrr.</i> sp. SS7-4 onto liposomes in the presence of Mg<sup>2+</sup>, HRPV-6 membrane fusion with the proteoliposomes could be readily observed. This was not the case with liposomes alone or with proteoliposomes carrying the S-layer extract from other haloarchaea, such as <i>Haloferax volcanii</i>. (4) Conclusions: The S-layer extract from the host, <i>Hrr.</i> sp. SS7-4, corresponds to the physiological fusion trigger of HRPV-6.


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