Cellular Proteins Associated with the Interior and Exterior of Vesicular Stomatitis Virus Virions

Abstract

Virus particles (virions) often contain not only virus-encoded but also host-encoded proteins. Some of these host proteins are enclosed within the virion structure, while others, in the case of enveloped viruses, are embedded in the host-derived membrane. While many of these host protein incorporations are likely accidental, some may play a role in virus infectivity, replication and/or immunoreactivity in the next host. Host protein incorporations may be especially important in therapeutic applications where large numbers of virus particles are administered. Vesicular stomatitis virus (VSV) is the prototypic rhabdovirus and a candidate vaccine, gene therapy and oncolytic vector. Using mass spectrometry, we previously examined cell type dependent host protein content of VSV virions using intact (“whole”) virions purified from three cell lines originating from different species. Here we aimed to determine the localization of host proteins within the VSV virions by analyzing: i) whole VSV virions; and ii) whole VSV virions treated with Proteinase K to remove all proteins outside the viral envelope. A total of 257 proteins were identified, with 181 identified in whole virions and 183 identified in Proteinase K treated virions. Most of these proteins have not been previously shown to be associated with VSV. Functional enrichment analysis indicated the most overrepresented categories were proteins associated with vesicles, vesicle-mediated transport and protein localization. Using western blotting, the presence of several host proteins, including some not previously shown in association with VSV (such as Yes1, Prl1 and Ddx3y), was confirmed and their relative quantities in various virion fractions determined. Our study provides a valuable inventory of virion-associated host proteins for further investigation of their roles in the replication cycle, pathogenesis and immunoreactivity of VSV.

Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication

“…It is well established that retroviruses package host lysyl-tRNA synthetase (LysRS) by contact with Gag, which targets tRNA to prime transcription to DNA upon infection of a new host ( 6 ). Host factors have also been previously observed in virus particles, such as influenza virus ( 7 ), vesicular stomatitis virus ( 8 ), vaccinia virus ( 9 ), and herpes simplex virus ( 10 ) (reviewed in reference 11 ).…”

Experimental Evolution Generates Novel Oncolytic Vesicular Stomatitis Viruses with Improved Replication in Virus-Resistant Pancreatic Cancer Cells

“…The recombinant viruses VSV-p53wt and VSV-p53-CC were previously engineered using the VSV-ΔM51 backbone and were previously described in detail (37). Purified virus was obtained exactly as described previously (105). Plaque-isolated viruses were obtained by isolating individual viral plaques, which were then amplified on BHK-21 cells.…”

CD59 association with infectious bronchitis virus particles protects against antibody-dependent complement-mediated lysis

“…Evolution has forced viruses to develop diverse mechanisms to avoid or delay destruction by the complement system [6]. To our knowledge, this article is the first to report that CD59 proteins are associated with IBV virions, although parainfluenza virus 5, mumps virus, vesicular stomatitis virus and NDV have been found to be associated with complement regulators through their envelopes [23][24][25][26][27]. Proteomic analysis of IBV particles has demonstrated that dozens of host proteins are present in the purified IBV virions, but CD59 was not detected [22,28].…”