Dataset Information

Time-course phosphoproteome response during adenovirus type 2 infection

ABSTRACT: Time-course systematic and quantitative analysis combining TiO2 enrichment and SILAC to investigate changes in the phosphoproteome of Ad2 and IMR-90 cells after adenovirus infection

INSTRUMENT(S): Q Exactive

ORGANISM(S): Human Adenovirus C Serotype 2 (hadv-2) (human Adenovirus 2) Homo Sapiens (human)

TISSUE(S): Fibroblast Of Lung, Cell Culture

DISEASE(S): Viral Infectious Disease

SUBMITTER: Sara Lind

LAB HEAD: Sara Lind

PROVIDER: PXD009562 | Pride | 2020-07-03

REPOSITORIES: Pride

ACCESS DATA

Publications

Phosphorylation Time-Course Study of the Response during Adenovirus Type 2 Infection.

Valdés Alberto A Zhao Hongxing H Pettersson Ulf U Lind Sara Bergström SB

Proteomics 20200225 7

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 (TiO<sub>2</sub> ) particle enrichment and SILAC-MS. Quantitative data from 1552 phosphorylated sites clustered the highly altered phosphorylated sites to the signaling by rho famil ...[more]

PMID: 32032466

Similar Datasets

Project description:To combat virus infections, which are major human killers, a deeper understanding of how viruses reprogram their hosts to create optimal production of progeny is needed. Most knowledge on the regulation of cellular gene expression during adenovirus infection is derived from studies of mRNA expression. Here, we investigated the changes in cellular protein expression during the late phase of adenovirus type 2 (Ad2) infection of primary human cells by stable isotope labeling in cell culture (SILAC) with subsequent liquid chromatography-high resolution tandem mass spectrometric (LC-MS/MS) analysis using a Q-Exactive Orbitrap instrument. Cells were in-depth evaluated 24 and 36 hours post infection (hpi) and two biological replicates were investigated using swapped labeling. In total, 2648 and 2394 proteins were quantified at 24 and 36 hpi, respectively. Among them, 659 and 645 were deregulated more than 1.6-fold at the two time points, and approximately 80% of them had sustained regulation from 24 to 36 hpi. For the deregulated genes the change in protein expression was compared with that of RNA expression using cDNA sequencing data from the same time points of infection. The comparison revealed a surprisingly low correlation (r = 0.3). Consistency between changes in RNA and protein expression was observed for approximately 30% of the genes, whereas a majority showed either divergent or opposite expression profiles. The deregulated proteins were subjected to pathway and biological function analysis using bioinformatics tools. A surprising observation was that proteins related to carbohydrate metabolism were up-regulated at the protein level but unchanged at the RNA level whereas histone proteins were down-regulated at the protein level but up-regulated at the RNA level. Our results show that regulation of cellular gene expression in the response to viral infection is more complex than hitherto believed and several examples of posttranscriptional regulation of gene expression were observed. The results contribute to our understanding of host cell gene regulation during infection at a deeper level.

Project description:The aquatic orthomyxovirus infectious salmon anemia virus (ISAV) is an important pathogen for salmonid aquaculture, however little is known about protective and pathological host responses to infection. We have investigated intracellular responses during cytopathic ISAV infection in the macrophage-like Atlantic salmon kidney (ASK) cell line by microarray analysis (1.8K SFA2.0 immunochip) and a functional assay for glutathione. Gene transcription changed rapidly and consistently with time and with minor differences between two virus isolates. While several pro-inflammatory and antiviral immune genes were induced, genes involved in cell signaling and integrity were down-regulated, suggesting isolation of infected cells from cell-to-cell interaction and responses to external signals. Differential expression of genes regulating cell cycle and apoptosis implied opposite cues from host cell and virus. This was in pace with massive down-regulation of genes involved in biosynthesis and processing of nucleotides and nucleic acids. Significant down-regulation of several genes involved in metabolism of reactive oxygen species suggested increased oxidative stress, which was confirmed by a functional assay showing reduced levels of glutathione during infection. Testing of expression data against a microarray database containing diverse experiments revealed candidate marker genes for ISAV infection. Our findings provide novel insight into cellular host responses and determinants for acute cytopathic ISAV infection. Keywords: Infectious salmon anemia virus (ISAV); Host response; Molecular pathology; Microarray; Macrophage; Oxidative stress ASK cells infected with ISAV isolate 2 and 4 and mock-infected were cultured in triplicates and total RNA pooled from each at the following time-points post-infection; 1, 3 and 5 days. For microarray analysis, single-slide hybridisation was applied between infected RNA (Cy5 labelled) and control RNA (Cy3 labelled) for each time-point. Results were verified by real-time qPCR.

Dataset Information

Time-course phosphoproteome response during adenovirus type 2 infection

Publications

Phosphorylation Time-Course Study of the Response during Adenovirus Type 2 Infection.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure