Taken together, accumulating evidence suggests that p23 is a viroporin that might form calcium-conducting channels in the ER membranes. Finally, flow cytometry studies indicate slightly elevated calcium concentrations in the cytoplasm of p23-transfected cells. We investigated changes to the global transcriptome and proteome that occurred in infected rabbit liver tissue and observed changes to the misfolded protein response, calcium signaling, and the regulation of the endoplasmic reticulum (ER) network. Furthermore, we identified cellular interactors of p23 using stable isotope labeling with amino acids in cell culture (SILAC)-based proteomics heat shock proteins Hsp70 and 110 interact with p23 in transfected cells, suggesting that they ‘chaperone’ p23 proteins before their integration into cellular membranes. Site-directed mutagenesis revealed the importance of an N-terminal cysteine for dimerization. Here, we report on the mechanistic details of p23 oligomerization. Furthermore, the positional homologs of p23 in other caliciviruses have been shown to possess viroporin activity. Instead, a range of in vitro and in silico models have been used to study p23, revealing that p23 oligomerizes, accumulates in the cytoplasm, and possesses a conserved C-terminal region with two amphipathic helices. The exact function(s) of the lagovirus non-structural protein p23 is unknown as robust cell culture systems for the Rabbit haemorrhagic disease virus (RHDV) and other lagoviruses have not been established. 10Land and Water, Commonwealth Scientific and Industrial Research Organisation, Canberra, ACT, Australia.9Australian Centre for Disease Preparedness, Commonwealth Scientific and Industrial Research Organisation, Geelong, VIC, Australia.8Research School of Chemistry, Australian National University, Acton, ACT, Australia.7Agriculture and Food, Commonwealth Scientific and Industrial Research Organisation, Canberra, ACT, Australia.6Imaging and Cytometry Facility, John Curtin School of Medical Research, Acton, ACT, Australia.5Centre for Proteome Research, Department of Biochemistry & Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, United Kingdom.4RSB/RSC Joint Mass Spectrometry Facility, Research School of Chemistry, Australian National University, Acton, ACT, Australia.3Centre for Invasive Species Solutions, Canberra, ACT, Australia.2Faculty of Science and Technology, University of Canberra, Canberra, ACT, Australia.1Health and Biosecurity, Commonwealth Scientific and Industrial Research Organisation, Canberra, ACT, Australia.Hall 1,3, Tanja Strive 1,3 and Michael Frese 1,2* Carroll 4, Joseph Boileau 4, Edward Emmott 5, Maria Jenckel 1, Harpreet Vohra 6, Vivien Rolland 7, Philip Hands 7, Junna Hayashi 8, Matthew J. The CPTR core has established over 200 targets identifying the following key wathways:Īpoptosis/Cell Death, Cell cycle and checkpoint control, Cellular metabolism, Chromatin Regulation/Epigenetics, DNA damage and repair, Gene regulation and DNA repair, JAK/STAT signaling, MAP Kinase signaling, NFκB signaling, PI3K/AKT/mTOR signaling, Protein Kinase C signaling, Receptor tyrosine Kinase signaling, Rho signaling, RNA regulation, TGF-β/SMAD signaling, Transcription regulation, Ubiquitin-proteasome, Wnt Signaling etc.Elena Smertina 1,2,3, Adam J. Assays identify critical pathways and molecular functioning mechanisms involved in the behavior of newly developed cell lines, PDXs, and clinical tumor samples. The assays provide custom pathway network profiling based on disease, drug target(s) etc. Straight forward assay standardization across multiple testing sites Simple Western Assays.Multiplex analysis with fast assay turn-around time.Nanogram (ng) level protein loading, capillary platform allows quantitative proteomic analysis in extremely small and precious samples, such as stem cells, primary cells, fine needle aspirates, other patient specimens.Precise and accurate measurement, digital data quantitation, good assay sensitivity, reproducibility and dynamic range.Have been applied for quantitative proteomic analysis in both discovery research and clinical practice.ğully automated and robust, provides bioanalytical labs with reproducibility and sensitivity in western blotting methods.Ğmploys high-resolution MW (size-based) or isoelectric-focusing (IEF, charge-based) separation, followed by target-specific immunoprobing to profile proteins and respective post-translational modification isoforms.
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