Analysis of the Differential Host Cell Nuclear Proteome Induced by Attenuated and Virulent Hemorrhagic Arenavirus Infection

Abstract

Arenaviruses are important emerging pathogens and include a number of hemorrhagic fever viruses classified as NIAID category A priority pathogens and CDC potential biothreat agents. Infection of guinea pigs with the New World arenavirus Pichindé virus (PICV) has been used as a biosafety level 2 model for the Lassa virus. Despite continuing research, little is known about the molecular basis of pathogenesis, and this has hindered the design of novel antiviral therapeutics. Modulation of the host response is a potential strategy for the treatment of infectious diseases. We have previously investigated the global host response to attenuated and lethal arenavirus infections by using high-throughput immunoblotting and kinomics approaches. In this report, we describe the differential nuclear proteomes of a murine cell line induced by mock infection and infection with attenuated and lethal variants of PICV, investigated by using two-dimensional gel electrophoresis. Spot identification using tandem mass spectrometry revealed the involvement of a number of proteins that regulate inflammation via potential modulation of NF-B activity and of several heterogeneous nuclear ribonuclear proteins. Pathway analysis revealed a potential role for transcription factor XBP-1, a transcription factor involved in major histocompatibility complex II (MHC-II) expression; differential DNA-binding activity was revealed by electrophoretic mobility shift assay, and differences in surface MHC-II expression were seen following PICV infection. These data are consistent with the results of several previous studies and highlight potential differences between transcriptional and translational regulation. This study provides a number of differentially expressed targets for further research and suggests that key events in pathogenesis may be established early in infection.

Differential effect of acute and persistent Junín virus infections on the nucleo-cytoplasmic trafficking and expression of heterogeneous nuclear ribonucleoproteins type A and B

“…2007; Cammas et al, 2007;Kim et al, 2007;Lin et al, 2008Lin et al, , 2009Gui et al, 2010). A previous report showed that the arenavirus Pichinde virus causes a transient cytoplasmic translocation of hnRNP A1 (Bowick et al, 2009). Here, we demonstrated that acute JUNV infection induces the cytoplasmic accumulation of hnRNP A1 in most infected cells but, in accordance with previous results obtained for hepatitis C virus-infected cells (Kim et al, 2007), some infected cells exhibited a nuclear hnRNP A1 pattern of distribution.…”

Attenuated and Lethal Variants of Pichindé Virus Induce Differential Patterns of NF-κB Activation Suggesting a Potential Target for Novel Therapeutics

“…Pichindé virus (PICV) causes a similar disease in guinea pigs to Lassa fever in humans (18). By using two variants of PICV-P2, which causes a mild infection, and P18, which causes a lethal disease (1), we can begin to delineate the cellular responses that lead to a protective immune response, and those that lead to a severe hemorrhagic disease (3)(4)(5).…”

A Systems Biology Starter Kit for Arenaviruses

“…Data pertaining to the arenavirus-host interplay was compiled from published studies on arenavirus-host protein-protein interactions [8,9,10,11,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62], viral requirements for specific host factors [63,64,65,66,67,68,69,70,71,72,73,74,75,76,77,78,79,80,81,82,83], viral inhibition of host proteins [84,85,86], kinomics and proteomics studies of cellular changes in response to infection [12,13,14,15,87,88,89,90,91,92,93,94,95,96,97,98,99,100,101,102,103,…”

“…The node with the highest connectivity value was found to be p53, a master regulator of cellular processes, whose steady-state levels are decreased during arenavirus infection [15], although no mechanism for this observation has yet been identified. Other hubs, which have already been identified as arenaviral host targets, include nucleophosmin [15], involved in ribosome biogenesis, and cytoskeletal elements such as vimentin [15], tubulin [70,71,72,77] and actin [13,70,71,72]. Amongst hub proteins that have not been previously identified as viral host targets, we found general adapters in signal transduction pathways such as YWHAZ or SHC1, as well as the master regulator AKT1 kinase, which is involved in many cellular processes such as metabolism, proliferation, cell survival and growth, and angiogenesis.…”

“…Additionally, certain large-scale analyses of host proteomics [12,13,14], kinomics (study of phosphorylation changes) [15], transcriptomics [16,17,18], and metabolomics (changes in levels of small molecules) [19] have been undertaken to investigate the consequences of arenavirus infections. However, any resulting biological networks that were created only included the results of these individual studies.…”