Virus innexin expression in insect cells disrupts cell membrane potential and pH Free

Abstract

Certain parasitoid wasps are associated with Polydnaviruses, symbiotic viruses that encode virulence factors which are essential to successful parasitization by the wasp of a caterpillar host. Members of one group of Polydnaviruses, the Ichnoviruses, encode a multi-gene family known as Vinnexins. Vinnexins are homologues of insect gap junction genes, and form functional gap junctions that may affect host cell physiology. However, the role of Vinnexins in host pathology and the mechanism by which these affect their caterpillar host are largely unknown. In this article, we generated recombinant baculoviruses to express vinnexins in Spodoptera frugiperda (Sf9) cells. To measure cell physiological changes caused by Vinnexins, cells were probed with a membrane potential-sensitive probe, DiBac4(3), and a pH indicator, carboxyfluorescein diacetate (CFDA). In addition, we utilized carbenoxolone and ouabain, respectively, to probe the role of gap junctions and hemi-channels, and Na+/K+-ATPase in establishing membrane potential in studied cells. Our results indicate that Vinnexins induce cell membrane depolarization and cytoplasmic alkalization to a degree specific to each tested Vinnexin, and that neither Vinnexin hemi-channels nor Na+/K+-ATPase appear to underlie these effects directly. These results hint that members of the Vinnexin protein family may affect host bio-electrical phenomena to disrupt host cell physiology, and that the individual proteins of the family may differentially affect host physiology.

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2018-08-09
2024-03-29
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