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Published on: September 16, 2016
Phosphorylation Time-Course Study of the Response during Adenovirus Type 2 Infection
Alberto Valdés1,2, Hongxing Zhao3, Ulf Pettersson3
1Section of Analytical Chemistry, Department of Chemistry-BMC, Uppsala University, Uppsala, 751 24, Sweden.
Abstract:
PTMs such as phosphorylations are usually involved in signal transduction pathways. To investigate the temporal dynamics of phosphoproteome changes upon viral infection, a model system of IMR-90 cells infected with human adenovirus type 2 (Ad2) is used in a time-course quantitative analysis combining titanium dioxide (TiO2 ) particle enrichment and SILAC-MS. Quantitative data from 1552 phosphorylated sites clustered the highly altered phosphorylated sites to the signaling by rho family GTPases, the actin cytoskeleton signaling, and the cAMP-dependent protein kinase A signaling pathways. Their activation is especially pronounced at early time post-infection. Changes of several phosphorylated sites involved in the glycolysis pathway, related to the activation of the Warburg effect, point at virus-induced energy production. For Ad2 proteins, 32 novel phosphorylation sites are identified and as many as 52 phosphorylated sites on 17 different Ad2 proteins are quantified, most of them at late time post-infection. Kinase predictions highlighted activation of PKA, CDK1/2, MAPK, and CKII. Overlaps of kinase motif sequences for viral and human proteins are observed, stressing the importance of phosphorylation during Ad2 infection.
Insights
Viral infection alters cell signaling and energy production by changing protein phosphorylation. This study reveals key pathways and viral phosphorylation sites during adenovirus infection in human cells.
Area of Science:
- Cellular Biology
- Virology
- Biochemistry
Background:
- Post-translational modifications (PTMs), particularly phosphorylation, are crucial for cellular signal transduction.
- Understanding how viral infections impact host cell phosphoproteome dynamics is essential for deciphering virus-host interactions.
Purpose of the Study:
- To investigate the temporal dynamics of phosphoproteome changes in IMR-90 cells following human adenovirus type 2 (Ad2) infection.
- To identify host and viral protein phosphorylation sites and their associated signaling pathways.
Main Methods:
- Time-course quantitative analysis using Stable Isotope Labeling by Amino acids in Cell culture (SILAC)-Mass Spectrometry (MS).
- Enrichment of phosphopeptides using titanium dioxide (TiO2) particles.
- Bioinformatic analysis for pathway clustering and kinase prediction.
Main Results:
- 1552 phosphorylated sites were quantified, with significant alterations in Rho family GTPases, actin cytoskeleton, and cAMP-dependent protein kinase A signaling pathways early post-infection.
- Changes in glycolysis-related phosphorylation sites suggest virus-induced Warburg effect activation for energy production.
- 32 novel and 52 known phosphorylation sites on 17 Ad2 proteins were identified, predominantly late post-infection.
- Kinase predictions indicated activation of PKA, CDK1/2, MAPK, and CKII.
Conclusions:
- Ad2 infection profoundly impacts host cell signaling and metabolism through dynamic phosphoproteome alterations.
- Phosphorylation plays a critical role in both host response and viral protein function during Ad2 infection.
- Observed overlaps in kinase motifs between viral and host proteins highlight conserved phosphorylation mechanisms.
