Quantitative proteomic analysis of Murine Norovirus infected BV-2 cells
Ontology highlight
ABSTRACT:
INSTRUMENT(S): LTQ Orbitrap Velos
ORGANISM(S): Mus Musculus (mouse)
TISSUE(S): Cell Culture, Microglial Cell
DISEASE(S): Norovirus Gastroenteritis
SUBMITTER: 
Edward Emmott
LAB HEAD: Ian Goodfellow
PROVIDER: PXD004015 | Pride | 2016-06-28
REPOSITORIES: Pride
Similar Datasets
Project description:SILAC-labelling analysis of murine BV-2 cells infected at high multiplicity of infection (10 TCID50/cell) and harvested at 4h or 9h post-infection. Represents the first quantitative proteomics analysis of norovirus-infected cells. A paired AP-MS dataset investigates the effects of MNV infection on eukaryotic initiation factor (eIF) complex formation with this dataset providing data on the overall abundance of eIF components in infected cells. This is a reanalysis of an earlier dataset (PXD004015) performed in Maxquant (v1.5.5.1).
2017-01-17 | PXD004984 | Pride
Project description:SILAC-labelling analysis of murine BV-2 cells infected at high multiplicity of infection (10 TCID50/cell) and harvested at 4h or 9h post-infection. Lysates were then suject to m7GTP-sepharose affinity purification to enrich for translation initiation factors. As norovirus translation uses a VPg protein covalently linked to the 5' of its RNAs for initiation factor recruitment it was hypothesised that norovirus infection could modify initiation factor complexes. A paired dataset investigates the effects of MNV infection on total protein levels in infected cells (PXD004015).
2016-07-07 | PXD004019 | Pride
Project description:SILAC-labelling analysis of murine BV-2 cells infected at high multiplicity of infection (10 TCID50/cell) and harvested at 4h or 9h post-infection. Lysates were then suject to m7GTP-sepharose affinity purification to enrich for translation initiation factors. As norovirus translation uses a VPg protein covalently linked to the 5' of its RNAs for initiation factor recruitment it was hypothesised that norovirus infection could modify initiation factor complexes. A paired dataset investigates the effects of MNV infection on total protein levels in infected cells. This dataset is a Maxquant reanalysis of data previously submitted under PXD004019
2017-01-17 | PXD004983 | Pride
Project description:Bluetongue virus causes infections in wild and domesticated ruminants, with the potential for causing significant morbidity, mortality and economic harm in both the developing and developed world. While vaccines have been developed, no treatment for acute infections exists. In this regard, a possible approach is to determine host-cell factors utilised by Bluetongue virus. We thus proceeded to characterize the phosphoproteome of BTV infected HeLa cells to elucidate the intracellular signalling pathways and identify critical host factors activated during Bluetongue virus infection.
2017-08-29 | PXD005550 | Pride
Project description:A hallmark of solid tumours is the development of hypoxic regions where rapidly growing transformed cells outstrip their blood supply. Tumour cells in these starved regions sense reduced levels of oxygen and switch on genes that help them to adapt, survive and continue growing. Since hypoxia is a key physiological difference between normal and cancer tissue, an opportunity exists to selectively kill tumour cells by exploiting the differences in protein expression between normal and hypoxic tumour tissue. However, a major challenge is to identify druggable hypoxia-induced proteins critical for tumour cell survival. This is especially important in cancers of unmet need where hypoxia gene signatures correlate with poor prognosis (for example, colorectal cancers). To identify new hypoxia-induced proteins, we performed SILAC-based proteomics on colorectal cancer cells exposed to normoxia and hypoxia. In this study, we identify a new hypoxia-activated GPCR signalling axis that enables colorectal tumour cells to survive the microenvironmental stress of hypoxia. Our findings uncover a previously unappreciated role for this GPCR-axis as a key regulator of the adaptive response to hypoxia and highlght an opportunity to exploit tumour-associated hypoxia for therapy.
2018-09-18 | PXD009971 | Pride
Project description:Cord blood stem cells are an attractive starting source for the production of red blood cells in vitro for therapy because of additional expansion potential compared to adult peripheral blood progenitors, and cord blood banks usually being more representative of national populations than blood donors. Consequently it is important to establish how similar cord RBCs are to adult cells. In this study we used Multiplex TMTs combined with nanoLC-MS/MS to compare the proteome of adult and cord RBCs and reticulocytes. 2838 unique proteins were identified, providing the most comprehensive compendium of RBC proteins to date. Using stringent criteria 1674 proteins were quantified, and only a small number differed in amount between adult and cord RBC. We focussed on proteins critical for RBC function. Of these only the expected differences in globin subunits, along with higher levels of carbonic anhydrase 1 & 2 and aquaporin-1 in adult RBCs would be expected to have a phenotypic effect since they are associated with the differences in gaseous exchange between adults and neonates. Since the RBC and reticulocyte samples used were autologous, we catalogue the change in proteome following reticulocyte maturation. The majority of proteins (>60% of the 1671 quantified) reduced in abundance between 2 and 100-fold following maturation. However, ~5% were at a higher level in RBCs, localised almost exclusively to cell membranes, in keeping with the known clearance of intracellular recycling pools during reticulocyte maturation. Overall, these data suggest that with respect to the proteome there is no barrier to the use of cord progenitors for the in vitro generation of RBCs for transfusion to adults other than the expression of fetal not adult haemoglobin.
2016-03-31 | PXD003276 | Pride
Project description:Total proteome and phospho proteome of two human cell lines infected with the ChAdOx1 nCoV-19 virus over 72 hours
2021-01-11 | PXD023286 | Pride
Project description:Immunoprecipitates were prepared from test Er(a+b-) donor RBCs and negative control Er(a-b+) (P1) RBCs using anti-Era plasma from P3 (5:1 ratio of plasma to cells) and proteomic analysis was performed.
2024-01-26 | PXD035013 | Pride
Project description:Platelets are small, anucleate cells that play an essential role in haemostasis, but can contribute critically to the pathogenesis of cardiovascular disease. Platelets express all four Class I Phosphoinositide 3-kinase (PI3K) isoforms, with previous work revealing important roles for these enzymes in regulating platelet priming, activation and stable thrombus formation. Despite this, detailed mechanistic understanding of how these lipid kinases orchestrate these roles in platelets is limited. Class I PI3Ks predominantly regulate cell function through their catalytic product, the signalling phospholipid phosphatidylinositol-3,4,5-trisphosphate (PIP3), which coordinates the localisation and/or activity of a diverse range of binding proteins. The complete repertoire of these Class I PI3K effectors in platelets remains unknown, hampering current understanding of Class I PI3K-mediated regulation of platelet function. We therefore conducted a global, unbiased screen for PIP3-binding proteins in human platelets using affinity capture coupled to high resolution proteomic mass spectrometry. Thus, we present the first in depth analysis of the PIP3 signalosome of human platelets, revealing an extensive PIP3 interactome, including many proteins previously uncharacterised in this cell type. This work provides important new insights into how Class I PI3K shapes human platelet function.
2017-06-06 | PXD003777 | Pride
Project description:We are here presenting a new paracrine induction of RNA granules by viruses. Infection by viruses imposes major stress on the host cell. In response to this stress, infected cells can induce several defence mechanisms, which include the activation of stress response pathways and the innate immune response. These often result in an inhibition of translation culminating in the assembly of cytoplasmic granules called stress granules (SGs). SGs assembly follows from liquid phase separation of aggregation-prone proteins such G3BP1 and TIA-1, leading to the sequestration of mRNAs. Because this threatens viral gene expression, viruses need to evade these stress response pathways to propagate. Using feline calicivirus (FCV), surrogate for norovirus, the main virus responsible for gastroenteritis outbreaks worldwide, we previously showed that FCV impairs SGs assembly by cleaving the scaffold protein G3BP1. Interestingly, we observed that uninfected bystander cells assembled G3BP1 granules, suggesting a paracrine response trigged by the infection. We now present evidence that virus-free supernatant generated from infected cells can induce the formation of RNA granules. We have characterised the dynamic of the granules assembly via confocal microscopy. Moreover, we provide an understanding of paracrine granules function and specificity through their affinity purification followed by proteomics and RNAseq analysis of their proteins and mRNAs content. This helps to define rules of assembly and novel functions for paracrine granules highlighting fundamental differences with canonical stress granules.
2022-03-14 | PXD021881 | Pride