FROM NIPAH VIRUS HOST INFECTION TO THE PRODUCTION OF NEW VIRAL PARTICLES
|Johnston, Gunner Pete
|Whittaker, Gary R.
|Collins, Ruth N.
|Vogt, Volker M.
|Supplemental file(s) description: Tables 4.1 and 4.2
|Nipah virus (NiV) belongs to the viral family Paramyxoviridae and is a pathogen of significant threat to global human health and agriculture. The absence of approved vaccines or treatments for NiV, severe human mortality reaching 100% rates in recent outbreaks, and the identification of at least twenty related viruses, underlines the urgency with which NiV and related pathogens should be studied. Despite prior reports that the NiV matrix (M) protein is the only viral protein significantly involved in particle formation and budding, we report that the NiV fusion (F) protein supports budding by enriching incorporation of the NiV attachment (G) protein into particles. Further, we report that this novel function of NiV F is dependent on motifs within its cytoplasmic tail, indicating potential dependence on cellular machinery. These findings were expanded upon with isolation of viral particles produced from different combinations of F, G, and M and subsequent proteomic analysis of such particles. These analyses and validation identified F-driven budding as most associated with cellular machinery, particularly vesicular trafficking and the actin cytoskeleton. Cellular factors involved in regulating endocytosis and recycling as well as ESCRTs were highly modulatory of F budding whether F was expressed alone or co-expressed with G and M. Finally, by broadening our use of high-throughput and bioinformatic technologies to “live” NiV infection at bio-safety level 4 conditions, we describe a novel study offering a comprehensive overview of infection. This multi-omics approach, combining transcriptomics, proteomics, metabolomics, and lipidomics indicated drastic changes to human cells affecting major processes including immune response, metabolism, and gene expression. Interestingly, we report substantial decreases in the abundance of proteins involved in translation, however, proteins associated with numerous, specific processes such as RNA processing exhibited dramatic enrichment. Interestingly, we also report major discrepancies between transcriptomics and proteomics, suggesting probable regulation of post-transcriptional protein expression during infection. Metabolic and lipidomic profiles supported shunting of glucose into nucleotide synthesis, the use of glutamine in anaplerosis, and the use of triglyceride catabolism to support cellular function. These findings have not been reported before for a henipavirus infection, begging further studies into their importance to infection.
|Attribution-NonCommercial-NoDerivatives 4.0 International
|FROM NIPAH VIRUS HOST INFECTION TO THE PRODUCTION OF NEW VIRAL PARTICLES
|dissertation or thesis
|Biomedical and Biological Sciences
|Doctor of Philosophy
|Ph.D., Biomedical and Biological Sciences