Project description:The purpose of this experiment was to evaluate the human host cellular response to wild-type Human coronavirus strain 229E (HCoV-229E) infection. Sample data was obtained for mock and infected (MOI 5) immortalized human lung epithelial cells (A549) and immortalized human lung fibroblasts cells (MRC5) processed for RNA sequencing (RNA-Seq) expression analysis.
Project description:The purpose of this experiment was to evaluate how wild-type Human coronavirus strain 229E (HCoV-229E) infection alters chromatin accessiblity in infected but not mock-infected samples.
Project description:Human coronaviruses (HCoVs) cause mild to severe respiratory infection. Most of the common cold illnesses are caused by one of four HCoVs, namely HCoV-229E, HCoV-NL63, HCoV-HKU1 and HCoV-OC43. Several studies have applied global transcriptomic methods to understand host responses to HCoV infection, with most studies focusing on the pandemic severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome CoV (MERS-CoV) and the newly emerging SARS-CoV-2. In this study, Next Generation Sequencing was used to gain new insights into cellular transcriptomic changes elicited by alphacoronavirus HCoV-229E. HCoV-229E-infected MRC5 cells showed marked downregulation of superpathway of cholesterol biosynthesis and eIF2 signaling pathways. Moreover, upregulation of cyclins, cell cycle control of chromosomal replication, and role of BRCA1 in DNA damage response, alongside downregulation of the cell cycle G1/S checkpoint, suggest that HCoV-229E favors S phase for viral infection. Intriguingly, more than 80% of key factors of cell innate immunity, interferon-stimulated genes (ISGs) and other transcripts of early antiviral response genes were downregulated early in HCoV-229E infection. This study will enhance our understanding of commonly circulating HCoVs and hopefully provide critical information about still-emerging coronaviruses.
Project description:The aim of the experiment is to identify changes in host gene expression upon infection of HuH7 human hepatoma cells with Human corona virus-229E (HCoV-229E) compared to mock-infected cells. All experiments were performed at 33°C which is required for sufficient viral entry/replication.
Project description:The purpose of this experiment was to evaluate the human host cellular response to wild-type Human coronavirus strain 229E (HCoV-229E) infection. Sample data was obtained for mock and infected (MOI 3) primary human airway epithelial cells grown in air-liquid interface conditions, processed for RNA sequencing (RNA-Seq) expression analysis.
Project description:In A549 cells, infection with human Coronavirus 229E spreads lateraly through the cell layer. This offers the opportunity to compare the gene expression patterns of infected cells with those of nearby unifected cells and to analyze paracrine effects occuring during infection. The aim of the experiment is to identify and compare changes in host gene expression of infected cells, cells immediately adjacent to infected cells and more distant cells with uninfected cells. To separate the different cell population, a Leica LMD6000 system was used. Individual cells were stained for the expression of the viral N protein or for DNA polymerase II by a shortened immunfluorescence protocol. After drying, the cell populations were visualized by fluorescence microscopy and separated by microdissection, collected in tubes and RNA was isolated on a small scale by column purification.
Project description:TMT18-labeled proteome-wide profiling of post-translational modifications in mock-infected and infected human lung epithelial cells (A549). Human lung epithelial (A549) cells were infected and mock-infected with HCoV-229E. Cells were harvested at 8, 16, and 24 hours post-infection. Extracted proteins were digested with LysC + Trypsin, then TMT18-labeled. Samples were split into 6 fractions and run on the Orbitrap Exploris 480. Data was searched with MS-GF+ using PNNL's DMS Processing pipeline.
Project description:TMT18-labeled proteome-wide profiling of post-translational modifications (acetylation) in mock-infected and infected human lung epithelial cells (A549). Human lung epithelial (A549) cells were infected and mock-infected with HCoV-229E. Cells were harvested at 8, 16, and 24 hours post-infection. Extracted proteins were digested with LysC + Trypsin, then TMT18-labeled and enriched for acetylation. Samples were split into 6 fractions and run on the Orbitrap Exploris 480. Data was searched with MS-GF+ using PNNL's DMS Processing pipeline.
Project description:TMT18-labeled proteome-wide profiling of post-translational modifications (phosphorylation) in mock-infected and infected human lung epithelial cells (A549). Human lung epithelial (A549) cells were infected and mock-infected with HCoV-229E. Cells were harvested at 8, 16, and 24 hours post-infection. Extracted proteins were digested with LysC + Trypsin, then TMT18-labeled and enriched for phosphorylation. Samples were split into 6 fractions and run on the Orbitrap Exploris 480. Data was searched with MS-GF+ using PNNL's DMS Processing pipeline.
