Proteomic analysis of mitochondria in respiratory epithelial cells infected with human respiratory syncytial virus and functional implications for virus and cell biology

Abstract

Objectives The aim of this study was to quantitatively characterise the mitochondrial proteome of airway epithelial cells infected with human respiratory syncytial virus (HRSV), a major cause of paediatric illness. Methods Quantitative proteomics, underpinned by stable isotope labelling with amino acids in cell culture, coupled to LC-MS/MS, was applied to mitochondrial fractions prepared from HRSV-infected and mock-infected cells 12 and 24 h postinfection. Datasets were analysed using ingenuity pathway analysis, and the results were validated and characterised using bioimaging, targeted inhibition and gene depletion. Key findings The data quantitatively indicated that antiviral signalling proteins converged on mitochondria during HRSV infection. The mitochondrial receptor protein Tom70 was found to act in an antiviral manner, while its chaperone, Hsp90, was confirmed to be a positive viral factor. Proteins associated with different organelles were also co-enriched in the mitochondrial fractions from HRSVinfected cells, suggesting that alterations in organelle dynamics and membrane associations occur during virus infection. Conclusions Protein and pathway-specific alterations occur to the mitochondrial proteome in a spatial and temporal manner during HRSV infection, suggesting that this organelle may have altered functions. These could be targeted as part of potential therapeutic strategies to disrupt virus biology.

Chaperoning the Mononegavirales: Current Knowledge and Future Directions

“…The chaperone dependence of numerous viral proteins from the Mononegavirales remains to be defined (e.g., SH, VP24, NS1, NS2, M2-1, M2-2, V, and C). Proteomics studies frequently identify chaperones as interacting partners [11,76,79,82,93,112,120], but many times they lack validation of the interaction and formal testing of the relevance of the identified chaperone in the context of infection. Even for well-studied interactions, such as that of the NC with Hsp70, or the L protein with Hsp90, the role of co-chaperones or upstream and downstream chaperone systems remains to be fully elucidated.…”

“…To date, the most druggable chaperone has been Hsp90. Work using various Hsp90 inhibitors has shown that non-toxic concentrations display antiviral activity against numerous Mononegavirales , including: the filovirus EBOV [110]; the paramyxoviruses PIV2 [88], MeV [86,111], MuV [80], and SV5 [88]; the pneumovirus RSV [62,79,87,112]; and the rhabdoviruses RABV [89] and VSV [88]. The successful inhibition of Hsp90 in humans for cancer treatment [26,43] highlights the feasibility of antiviral approaches that target chaperones.…”

Chaperoning the <em>Mononegavirales</em>: Current Knowledge and Future Directions

“…SH, VP24, NS1, NS2, M2-1, M2-2, V, C). Proteomics studies have described some interactions with chaperones for a subset of these [11,75,87], as well as for better studied viral proteins [67,70,112,119]. However, these lack both validation and formal testing of a role in the viral life cycle.…”

“…To date, the most druggable chaperone has been Hsp90. Work using various Hsp90 inhibitors has shown that non-toxic concentrations display antiviral activity against numerous Mononegavirales, including the filovirus EBOV [105]; the paramyxoviruses PIV2 [82], MeV [79,111], MuV [72], and SV5 [82]; the pneumovirus RSV [52,70,81,112]; and the rhabdoviruses RABV [83] and VSV [82]. The successful inhibition of Hsp90 in humans for the treatment of cancer [26,43] highlights the feasibility of antiviral approaches targeting chaperones.…”

Enhancing vaccine design strategies: applications for protein science, proteomics and adjuvants

“…[2] In another powerful application of proteomics, the paper by Munday et al explores how the syncytial viral infection of epithelial cells alters expression of mitochondrial proteins in a temporal manner. [3] While Tom was found to be protective, hsp90 supported epithelial cell infection by virus. [3] Such knowledge offers novel targets to explore in the development of anti-infective strategies.…”