title: Mass spectrometry reveals specific and global molecular transformations during viral infection.
abstract: Mass spectrometry analysis was used to target three different aspects of the viral infection process: the expression kinetics of viral proteins, changes in the expression levels of cellular proteins, and the changes in cellular metabolites in response to viral infection. The combination of these methods represents a new, more comprehensive approach to the study of viral infection revealing the complexity of these events within the infected cell. The proteins associated with measles virus (MV) infection of human HeLa cells were measured using a label-free approach. On the other hand, the regulation of cellular and Flock House Virus (FHV) proteins in response to FHV infection of Drosophila cells was monitored using stable isotope labeling. Three complementary techniques were used to monitor changes in viral protein expression in the cell and host protein expression. A total of 1500 host proteins was identified and quantified, of which over 200 proteins were either up- or down-regulated in response to viral infection, such as the up-regulation of the Drosophila apoptotic croquemort protein, and the down-regulation of proteins that inhibited cell death. These analyses also demonstrated the up-regulation of viral proteins functioning in replication, inhibition of RNA interference, viral assembly, and RNA encapsidation. Over 1000 unique metabolites were also observed with significant changes in over 30, such as the down-regulated cellular phospholipids possibly reflecting the initial events in cell death and viral release. Overall, the cellular transformation that occurs upon viral infection is a process involving hundreds of proteins and metabolites, many of which are structurally and functionally uncharacterized.
authors.affiliation: Department of Molecular Biology and The Center for Mass Spectrometry, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Eden P
authors.initials: EP
authors.lastname: Go
authors.forename: William R
authors.initials: WR
authors.lastname: Wikoff
authors.forename: Zhouxin
authors.initials: Z
authors.lastname: Shen
authors.forename: Grace
authors.initials: G
authors.lastname: O'Maille
authors.forename: Hirotoshi
authors.initials: H
authors.lastname: Morita
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Conrads
authors.forename: Anders
authors.initials: A
authors.lastname: Nordstrom
authors.forename: Sunia A
authors.initials: SA
authors.lastname: Trauger
authors.forename: Wilasinee
authors.initials: W
authors.lastname: Uritboonthai
authors.forename: David A
authors.initials: DA
authors.lastname: Lucas
authors.forename: King C
authors.initials: KC
authors.lastname: Chan
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Veenstra
authors.forename: Hanna
authors.initials: H
authors.lastname: Lewicki
authors.forename: Michael B
authors.initials: MB
authors.lastname: Oldstone
authors.forename: Anette
authors.initials: A
authors.lastname: Schneemann
authors.forename: Gary
authors.initials: G
authors.lastname: Siuzdak
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004331:Drosophila melanogaster; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008459:Measles virus / Q000378:metabolism*; D030381:Nodaviridae / Q000378:metabolism*; D010103:Oxygen Isotopes; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012327:RNA Virus Infections / Q000378:metabolism*
chemical_list: D010103:Oxygen Isotopes; D011506:Proteins