Comparative proteomic analyses of two reovirus T3D subtypes and comparison to T1L identifies multiple novel proteins in key cellular pathogenic pathways

Abstract

Viruses induce changes in the host to facilitate replication and evade the immune response. These changes are reflected by the host's proteome, including differences in protein abundance. Focusing on up and down regulated proteins after a virus infects the cell will lead to a characterization of the host response to infection, and may give insight into how viruses modulate proteins to evade host defense responses. We previously used SILAC to examine host proteomic changes in protein abundance in HEK293 cells infected with reovirus type 1, strain Lang (T1L). For the present study, we extended this analysis by determining cell protein alterations induced by two different reovirus subtypes, a less pathogenic type 3 Dearing (T3D(F)) isolate, and a more pathogenic isolate named T3D(C) that is presently in clinical trials as an anti-cancer oncolytic agent. This comparison of host proteome regulation showed that T3D(C) had a more marked effect on DNA replication proteins, recombination and repair, as well as immunological, apoptotic, and survival cell functions. We also identified several proteins not previously identified in any virus infection; branched chain amino-acid transaminase 2 (BCAT), paternally expressed 10 (PEG10), target of myb1 (TOM1), histone cluster 2 H4b (HIST2H4B) and tubulin beta 4B (TUBB4B).

Single Amino Acid Differences between Closely Related Reovirus T3D Lab Strains Alter Oncolytic Potency In Vitro and In Vivo

“…These were some of the earliest reported differences between laboratory reovirus strains. More recently, comparisons between T3D laboratory strains found that T3D C (from the Cashdollar laboratory) and T3D F/N (from the Nibert laboratory) exhibit different respiratory pathogeneses in vivo and in cell signaling in vitro (23,41). Surprisingly, however, similar in vitro replication kinetics were observed between T3D laboratory strains compared to date.…”

ENTPD1 (CD39) Expression Inhibits UVR-Induced DNA Damage Repair through Purinergic Signaling and Is Associated with Metastasis in Human Cutaneous Squamous Cell Carcinoma

“…TNFRSF10C, which is a known p53-regulated DNA damage-inducible gene, was upregulated with adenosine treatment (Sheikh et al, 1999). Interestingly, several genes involved with viral pathogenesis including PEG10, MX1, IFI44, and DDX60 were upregulated after treatment with adenosine (Berard et al, 2015, Kaczkowski et al, 2012. Finally, adenosine treatment induced the expression of several genes implicated in the interferon gene response (IRF9, MX1, IFIT1) (Gerner et al, 2019).…”

Synthesis and Translation of Viral mRNA in Reovirus-Infected Cells: Progress and Remaining Questions

“…But it is only as recently as 2007, that a T7 plasmid-based reverse genetics technique finally allowed to alter with relative ease the genome of reovirus and thus study the effect of predetermined changes on the viral multiplication cycle [19,20,21]. More recently, high-throughput approaches have also been used to extensively look at the effect of reovirus infection on either the cellular proteome [22,23,24,25] or transcriptome using microarrays [26,27,28,29,30,31] or more extensive RNA-seq approaches [32,33]. Altogether, these approaches are likely to shed new light on the complex interactions between the host cell and the virus, as discussed below.…”