Journal
JOURNAL OF PROTEOME RESEARCH
Volume 5, Issue 9, Pages 2405-2416Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr060215t
Keywords
virus; protein regulation; viral infection; metabolites; isotope labeling; mass spectrometry
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Funding
- NCI NIH HHS [N01-CO-12400] Funding Source: Medline
- NIAID NIH HHS [AI036222, R01 AI036222] Funding Source: Medline
- NIGMS NIH HHS [R01 GM055775, GM55775, R01 GM055775-02] Funding Source: Medline
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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.
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