Using CC strains to dissect the genetic regulation of the proteome
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Orbitrap Fusion Lumos
ORGANISM(S): Mus Musculus (mouse)
TISSUE(S): Liver
SUBMITTER: 
Tian Zhang
LAB HEAD: Steven Gygi
PROVIDER: PXD018886 | Pride | 2021-07-29
REPOSITORIES: Pride
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| 363771_a11149.mzIdentML | Mzid | |||
| 363772_a11148.mzIdentML | Mzid | |||
| 363773_a11147.mzIdentML | Mzid | |||
| 363774_a11146.mzIdentML | Mzid | |||
| 363775_a11145.mzIdentML | Mzid |
Items per page: 5 1 - 5 of 288 |
Similar Datasets
Project description:To examine the functional consequences of intratumor heterogeneity, subclonal populations (SCPs) were derived from the MDA-MB-468 triple-negative breast cancer cell line. Characterization of these SCPs revealed considerable variation in SCP tumor forming capacity with SCP #32 (SCP32) having an exceptional high tumor forming capacity. To investigate the proteomic differences between tumors of SCP32 and other SCPs, in TMT1, isobaric tandem mass tag (TMT) labeling combined with LC-MS/MS (TMT-MS) was performed on tumors of SCP03, SCP29 and SCP32 (n = 3 each) collected at 2 months post transplantation. In TMT2, isobaric tandem mass tag (TMT) labeling combined with LC-MS/MS (TMT-MS) was performed on tumors of the parental MDA-MB-468 cell line (n = 2), and both barcoded and non-barcoded versions SCP29 and SCP32 (two each) collected at 2 months post transplantation.
2022-02-16 | PXD022325 | Pride
Project description:Here we describe a new class of immunostimulatory short duplex RNAs that potently induce production of type I interferon (IFN-I), and particularly IFN-β, in a wide range ofhuman cell types via end-to-end dimerization, direct binding to RIG-I, and activation of the RIG-I/IRF3 pathway. These RNAs require a minimum of 20 base pairs, lack any sequence or structural characteristics of known immunostimulatory RNAs, and instead require a unique conserved sequence motif (sense strand: 5’-C, antisense strand: 3’-GGG) that mediates the self-assembly of end-to-end RNA dimers by Hoogsteen G-G base-pairing. The presence of terminal hydroxyl or monophosphate groups, blunt or overhanging ends, or terminal RNA or DNA bases also did not affect their ability to induce IFN-I production. Immune stimulation mediated by these duplex RNAs results in broad spectrum inhibition of infections by many respiratory viruses with pandemic potential, including SARS-CoV-2, SARS-CoV, MERS-CoV, and influenza A, as well as the common cold virus HCoV-NL63 in cell lines and in human Lung Chips that mimic organlevel lung pathophysiology. These novel immunostimulatory motifs potentially could be harnessed to create broad-spectrum antiviral therapeutics, but they should be avoided when designing siRNAs to minimize immunological side effects.
2022-09-01 | PXD027838 | Pride
Project description:This project tessts whether translation of all selenoproteins are affected by NRF2 downregualation in A549 and H1437 cells.
2020-10-22 | PXD018634 | Pride
Project description:Phosphorylation of proteins is a key step in the regulation of many cellular processes including activation of enzymes and signaling cascades. Phosphorylated peptides (phosphopeptides) can be detected and quantified by mass spectrometry. The abundance of a phosphopeptide is determined by both the abundance of its parent protein and the proportion of target sites that are phosphorylated. We quantified phosphopeptides, proteins, and transcripts in heart, liver, and kidney tissue samples from female and male pair from 58 strains of the Collaborative Cross strain panel. We mapped ~700 phosphorylation quantitative trait loci (phQTL) across the three tissues and applied genetic mediation analysis to identify causal drivers of phosphorylation. We identified kinases, phosphatases, cytokines, and other factors, including both known and potentially novel interactions between target proteins and genes that regulate site-specific phosphorylation. We observed coordination of phosphorylation across multiple sites within a protein and across proteins that form complexes. Our analysis highlights multiple targets of pyruvate dehydrogenase kinase 1 (PDK1), a regulator of mitochondrial function that shows reduced activity in the NZO/HILtJ mouse, a polygenic model of obesity and type 2 diabetes.
2023-02-02 | PXD032843 | Pride
Project description:Saccharomyces cerevisiae and Schizosaccharomyces pombe are the most commonly studied yeast model systems, yet comparisons of global proteome remodeling between these yeast species are scarce. Here, we profile the proteomes of S. cerevisiae and S. pombe cultured with either glucose or pyruvate as the sole carbon source to define common and distinctive alterations in the protein landscape across species. In addition, we developed an updated streamlined-tandem mass tag (SL-TMT) strategy that substitutes chemical-based precipitation with more versatile bead-based protein aggregation method (SP3) prior to enzymatic digestion and TMT labeling. Our new workflow, SL-SP3-TMT, allowed for near-complete proteome profiles in a single experiment for each species. The data revealed expected alterations in protein abundance, highlighted complete canonical biochemical pathways, and provided insight into previously uncharacterized proteins. The techniques used herein, namely SL-SP3-TMT, can be applied to virtually any experiment aiming to study remodeling of the proteome using a high-throughput, comprehensive, yet streamlined mass spectrometry-based strategy.
2021-05-11 | PXD014546 | Pride
Project description:Cells harbor two systems for fatty acid synthesis, one in the cytoplasm (catalyzed by fatty acid synthase, FASN) and one in the mitochondria (mtFAS). In contrast to FASN, mtFAS is poorly characterized, especially in higher eukaryotes, with the major product(s), metabolic roles, and cellular function(s) being essentially unknown. Here we show that hypomorphic mtFAS mutant mouse skeletal myoblast cell lines display a severe loss of electron transport chain (ETC) complexes and exhibit compensatory metabolic activities including reductive carboxylation. This effect on ETC complexes appears to be independent of protein lipoylation, the best characterized function of mtFAS, as mutants lacking lipoylation have an intact ETC. Finally, mtFAS impairment blocks the differentiation of skeletal myoblasts in vitro. Together, these data suggest that ETC activity in mammals is profoundly controlled by mtFAS function, thereby connecting anabolic fatty acid synthesis with the oxidation of carbon fuels.
2020-09-02 | PXD021034 | Pride
Project description:Toll-like Receptors (TLRs) are key mediators of immune responses to infection. Upon microbial detection, these receptors activate inflammatory signal transduction pathways that involve IκB kinases, mitogen activated protein kinases, ubiquitin ligases and other adaptor proteins. Current models suggest these signaling proteins operate within functionally distinct multiprotein complexes, which are all activated by a receptor-proximal complex known as the myddosome. The mechanisms that connect the effector protein complexes in the TLR pathways are undefined. To define TLR pathway activities, we engineered macrophages to enable proteomic analysis of the endogenous myddosome constituent MyD88 and its interacting proteins. Proteomics demonstrated that myddosomes contain proteins that act at all stages and regulate all effector responses of the TLR signaling pathways.
2024-07-04 | PXD047416 | Pride
Project description:Genetic background is a major driver of the phenotypic variability observed across pluripotent stem cells (PSCs), and studies addressing it have relied on transcript abundance as the primary molecular readout of cell state. However, little is known about how proteins, the functional units in the cell, vary across genetically diverse PSCs and how this relates to variation in other measures of gene output. Here we present the first comprehensive genetic study characterizing the pluripotent proteome using 190 unique mouse embryonic stem cell lines derived from highly heterogeneous Diversity Outbred mice. Moreover, we integrated the proteome with chromatin accessibility and transcript abundance in 163 cell lines with matching genotypes using multi-omics factor analysis to distinguish shared and unique drivers of variability across molecular layers. Our findings highlight the power of multi-omics data integration in revealing the distal impacts of genetic variation. We show that limitations in mapping of individual molecular traits may be overcome by utilizing data integration to consolidate the influence of genetic signals shared across molecular traits and increase detection power.
2023-05-10 | PXD033001 | Pride
Project description:Electron transport chain (ETC) defects occurring from mitochondrial disease mutations compromise ATP synthesis and render cells vulnerable to nutrient and oxidative stress conditions. This bioenergetic failure is thought to underlie pathologies associated with mitochondrial diseases. However, the precise metabolic processes resulting from a defective mitochondrial ETC that compromise cell viability under stress conditions are not entirely understood. We design a whole genome gain-of-function CRISPR activation screen using human mitochondrial disease complex I (CI) mutant cells to identify genes whose increased function rescue glucose restriction-induced cell death. The top hit of the screen is the cytosolic Malic Enzyme (ME1), that is sufficient to enable survival and proliferation of CI mutant cells under nutrient stress conditions. Unexpectedly, this metabolic rescue is independent of increased ATP synthesis through glycolysis or oxidative phosphorylation, but dependent on ME1-produced NADPH and glutathione (GSH). Survival upon nutrient stress or pentose phosphate pathway (PPP) inhibition depends on compensatory NADPH production through the mitochondrial one-carbon metabolism that is severely compromised in CI mutant cells. Importantly, this defective CI-dependent decrease in mitochondrial NADPH production pathway or genetic ablation of SHMT2 causes strong inflammatory cytokine signatures associated with redox dependent induction of ASK1 and activation of stress kinases p38 and JNK. These studies find that a major defect of CI deficiencies is decreased mitochondrial one-carbon NADPH production that is associated with increased inflammation and cell death.
2021-09-09 | PXD018839 | Pride
Project description:The Ubiquitin-Proteasome System (UPS) regulates many cellular processes in eukaryotic cells. Ubiquitylation by the UPS mainly directs proteins to proteasomal degradation, but it can also have non-degradative functions, such as regulating protein activity or localization. The small protein ubiquitin is conjugated to its substrates via a cascade of E1-E2-E3 enzymes. Dysregulation of the UPS has been implicated in the genesis and progression of many diseases, such as neurodegenerative diseases and cancer; thus, the UPS components are attractive targets for developing pharmaceutical drugs. E2s, or ubiquitin conjugating enzymes, are central players of the UPS. E2s function in tandem with specific ubiquitin ligases (E3s) to transfer ubiquitin to substrates. Here, we present the first proteome stability analysis of two closely related ubiquitin conjugating enzymes, UBC4 and UBC5, in S. cerevisiae. These two E2s are nearly identical, having 92% sequence identity and differing by only 11 amino acid residues. This dataset is of broad interest because higher eukaryotes express ubiquitin conjugating enzymes that are analogous to the yeast UBC4/5.
2022-10-13 | PXD037315 | Pride