Double-membrane vesicle of Porcine Reproductive and Respiratory Syndrome virus
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Porcine Reproductive and Respiratory Syndrome (PRRS) is a global disease which takes a significant toll on the pork industry and the welfare of pigs. The causative agent – PRRS virus (PRRSV) – is a single-stranded, positive-sense RNA virus of the Nidovirales order. In the process of replication, PRRSV induces the rearrangement of cellular membranes to form double-membrane vesicles (DMVs). These structures are thought to have a role in a) concentrating viral structures to increase their chances of interacting with one another, and b) preventing elements of the cellular immune response from detecting viral structures. Previous work has suggested that the DMV originates from the autophagy pathway – a highly-conserved mechanism for cells to recycle extraneous organelles and proteins during times of stress. Other work suggests that the DMV may be a co-opted EDEMosome – a recently-discovered vesicle which is involved in regulating the level of endoplasmic reticulum-associated degradation (ERAD). This thesis explores these possibilities – using immunofluorescent imaging as well as examining the proteomic and ribonucleic acid composition of the DMV as isolated by flow cytometry or separated from other organelles by density gradient – calling both candidate pathways into question and suggesting other candidate structures such as exosomal vesicles.