Browse
Submit Data
Databases
API
Help

Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

A balance in the delivery of ER components and the vacuolar proton pump determines phagosome acidification

ABSTRACT: As part of a study comparing phagosome composition in wild-type Dictyostelium discoideum with mutants in two cytoskeletal/motor proteins, we compared global gene expression and proteomes between the strains to test whether differences in mRNA and protein abundance were correlated with changes in phagosomal composition. abp1 and myoK mutants in both the Ax2 and DH1 backgrounds were compared with their respective wild-type parents.Both analyses converged and confirmed the phagosome analysis (supplemental Tables). Protein differences were markedly fewer in the myoK-null mutant than in the abp1 null lysate. Proteins of known function, differential in the myoK-null lysate, were involved in remodeling of the plasma membrane and early membrane trafficking. Proteins differential in the abp1-null lysate spanned various functions at all levels of cell metabolism. Most of the proteins differential in 2D-DIGE displayed similar mutant-to-wt ratios at the mRNA level in microarrays. A prominent example of overlap is the 1.4 fold overexpression (p ??? 0.016) of Vps5, both in proteomics and microarray analyses of abp1-null cells. Like half of the proteins involved in membrane trafficking that are differential in microarray analyses of abp1-null cells, Vps5 is putatively involved in endosome-to-Golgi trafficking as a retromer sorting nexin. Therefore, one might speculate that, in addition to direct effects on phagocytic uptake, absence of Abp1 has an impact on endosome-to-Golgi trafficking.

ORGANISM(S): Dictyostelium discoideum

SUBMITTER: Gareth Bloomfield

PROVIDER: E-MTAB-978 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Json Xml

Similar Datasets

Quantitative proteome for C.elegans by 3plex mTRAQ

Project description:Vesicle-mediated transport is a fundamental part of the secretory and endocytic pathways. In addition to their role in membrane protein trafficking, Arf-like protein subfamily of small GTPases also plays an important role in development, maintenance of Golgi structure and function. Proper protein trafficking is critical for cellular integrity and studies demonstrate that aberrant protein sorting leads to various diseases in many organisms including humans. However, to date, there is no report showing the role of Golgi apparatus and Golgi proteins in organismal longevity. Organisms dynamically reprogram their transcriptome and proteome as a response to internal and external changes, which alter physiological processes including aging. Compared to many transcriptomic studies that identified aging-regulatory genes, proteomic studies that identified aging-regulatory proteins are relatively scarce. By using a quantitative proteomic approach, we identified MON-2, an Arf-Gef protein implicated in Golgi-to-endosome trafficking, as a longevity-promoting protein. We found that MON-2 is essential for the long lifespan of various longevity mutants. Our results demonstrate that Golgi-to-endosome trafficking is an integral part of lifespan regulation.

2022-02-17 | PXD026411 | Pride

Profiling nucleo/cytoplasmic distribution of mRNAs identifies a role for the Golgi-to-ER trafficking in C9orf72 toxicity

Project description:Compelling evidence indicates that defects in nucleocytoplasmic transport contribute to the pathogenesis of amyotrophic lateral sclerosis (ALS). In particular, hexanucleotide (G4C2) repeat expansions in C9orf72, the most common cause of genetic ALS, have a widespread impact on the transport machinery that regulates the nucleocytoplasmic distribution of proteins and RNAs, thereby affecting their functions. We have previously reported that the expression of G4C2 hexanucleotide repeats in cultured human and mouse cells caused a marked accumulation of poly(A) mRNAs in cell nuclei, suggesting that mRNA trafficking is impaired by expanded C9orf72-ALS repeats. To further characterize the process, in this work we set out to systematically identify the specific mRNAs that are altered in their nucleocytoplasmic distribution in the presence of C9orf72-ALS repeats. Results from RNAseq profiling of nuclear and cytoplasmic RNA fractions obtained from cells expressing G4C231 repeats show a significant accumulation of mRNAs in the nuclei of G4C231 cells, further suggesting that nuclear retention of mRNAs is a major effect of C9orf72 expanded repeats expression. Interestingly, pathway analysis shows that mRNAs involved in ER-to-Golgi trafficking are particularly enriched among the identified mRNAs. Most importantly, functional studies in cultured cells and Drosophila indicate that the membrane trafficking route regulated by ARFGAP1, a GTPase-activating protein that associates with Golgi, is specifically affected by expanded C9orf72 repeats, as well as by C9orf72-related dipeptide repeat proteins (C9-DPRs), pointing to ER-to-Golgi vesicle-mediated transport as a target of C9orf72 toxicity.

2023-08-21 | GSE235448 | GEO

Expression analysis of pre-phenotype dt27J mouse dorsal root ganglia

Project description:Loss of function of dystonin cytoskeletal linker proteins causes neurodegeneration in the sensory ataxia, dystonia musculorum (dt). While much investigation has focused on understanding dt pathology, divergent functions of dystonin isoforms are still unknown. Here, we highlight a novel function of the dystonin-a2 isoform in mediating microtubule (MT) stability, golgi organization and flux through the secretory pathway. Using dystonin-null mice combined with isoform-specific loss of function analysis, we find dystonin-a2 is bound to MAP1B in the area surrounding the centrosome, where it maintains MT acetylation. In dt, the absence of the MAP1B-dystonin-A2 interaction results in a loss of MAP1B perinuclear localization, leading to MT deacetylation and instability. Deacetylated MTs lead to golgi fragmentation and prevent anterograde trafficking of motor proteins. Maintenance of MT acetylation through TSA administration or MAP1B overexperssion in vitro, mitigates the observed defect. These aberrations are apparent in pre-phenotype dorsal root ganglia (DRG) and primary sensory neurons, suggesting they are causal in the dt disorder. P4 dorsal root ganglia (DRG) tissue from 3 WT and 3 dt27J animals from 2 separate litters was subjected to RNA extraction and analyzed.

2012-02-23 | E-GEOD-30960 | biostudies-arrayexpress

Golgi-IP, a tool for multimodal analysis of Golgi molecular content

Project description:The Golgi is a membrane-bound organelle that is essential for protein and lipid biosynthesis. It represents a central trafficking hub that sorts proteins and lipids to various destinations or for secretion from the cell. The Golgi has emerged as a docking platform for cellular signaling pathways including LRRK2 kinase whose deregulation leads to Parkinson disease. Golgi dysfunction is associated with a broad spectrum of diseases including cancer, neurodegeneration, and cardiovascular diseases. To allow the study of the Golgi at high resolution, we report a rapid Golgi immunoprecipitation technique (Golgi-IP) to isolate intact Golgi mini-stacks for subsequent analysis of their content. By fusing the Golgi-resident protein TMEM115 to three tandem HA epitopes (GolgiTAG), we purified the Golgi using Golgi-IP with minimal contamination from other compartments. We then established an analysis pipeline using liquid chromatography coupled with mass spectrometry to characterize the human Golgi proteome, metabolome, and lipidome. Subcellular proteomics confirmed known Golgi proteins and identified proteins not previously associated with the Golgi. Metabolite profiling established the human Golgi metabolome and revealed the enrichment of uridine-diphosphate (UDP) sugars and their derivatives, which is consistent with their roles in protein and lipid glycosylation. Furthermore, targeted metabolomics validated SLC35A2 as the subcellular transporter for UDP-hexose. Finally, lipidomics analysis showed that phospholipids including phosphatidylcholine, phosphatidylinositol, and phosphatidylserine are the most abundant Golgi lipids and that glycosphingolipids are enriched in this compartment. Altogether, our work establishes a comprehensive molecular map of the human Golgi and provides a powerful method to study the Golgi with high precision in health and disease.

2023-06-27 | MTBLS6511 | MetaboLights

The interactome of the UapA transporter reveals putative new players in anterograde membrane cargo trafficking: UapA Interactome

Project description:Neosynthesized plasma membrane (PM) proteins co-translationally translocate to the ER, concentrate at regions called ER-exit sites (ERes) and pack into COPII secretory vesicles which are sorted to the early-Golgi through membrane fusion. Following Golgi maturation, membrane cargoes reach the late-Golgi, from where they exit in clathrin-coated vesicles destined to the PM, directly or through endosomes. Post-Golgi membrane cargo trafficking also involves the cytoskeleton and the exocyst. The Golgi-dependent secretory pathway is thought to be responsible for the trafficking of all major membrane proteins. However, our recent findings in Aspergillus nidulans showed that several plasma membrane cargoes, such as transporters and receptors, follow a sorting route that seems to bypass Golgi functioning. To gain insight on membrane trafficking and specifically Golgi-bypass, here we used proximity dependent biotinylation (PDB) coupled with data-independent acquisition mass spectrometry (DIA-MS) for identifying transient interactors of the UapA transporter.

2023-11-21 | PXD043934 | Pride

Genetic Control of Cellular Quiescence in S. pombe

Project description:Transition from proliferation to quiescence brings about extensive changes in cellular behavior and structure. However, genes critical for establishing and/or for maintaining quiescence are largely unknown. The fission yeast S. pombe is found as an excellent model for studying this problem, because it becomes quiescent under nitrogen starvation. Here we characterize 610 temperature-sensitive (ts) mutants, and identify 33 genes required for entry into and the maintenance of quiescence. These genes cover a broad range of cellular functions in the cytoplasm, membrane and the nucleus, encoding proteins for stress-responsive and cell cycle kinase signaling pathway, actin-bound and osmo-controlling endosome formation, RNA transcription, splicing and ribosome biogenesis, chromatin silencing, biosynthesis of lipid and ATP, cell wall and membrane morphogenesis, protein trafficking and vesicle fusion. We specifically highlight Fcp1, CTD phosphatase of RNA polymerase II, which differentially affects transcription of genes involved in quiescence and proliferation. We propose that the transcriptional role of Fcp1 is central to differentiate quiescence from proliferation. Keywords: cell cycle control, protein trafficking, cortex actin, RNA polymerase II, protein phosphatase

2009-04-30 | GSE14319 | GEO

Single-Cell Analysis Reveals Androgen Receptor Regulates ER-to-Golgi Trafficking with CREB3L2 To Drive Prostate Cancer Progression

Project description:Androgen receptor (AR) plays an essential role in normal prostate development and prostate cancer (PCa) progression. To understand the role of AR at the single-cell level, we performed single-cell transcriptome analysis on PCa cells stimulated with androgen and antiandrogen to reconstruct the dynamic and direct AR transcriptional network. Our work reveals that androgen stimulates the ER and Golgi stress response , promoting secreting protein trafficking, and inhibiting cell apoptosis. Moreover, we identify an ER-to-Golgi protein vesicle-mediated transport gene signature essential for maximal androgen-mediated ER-Golgi trafficking, cell proliferation, and association with PCa prognosis and progression. Notably, we show that AR collaborates with CREB3L2, XXX, to coordinately promote ER-Golgi trafficking of Golgi enzyme Mannosidase II and PCa cell survival. Finally, we show the inhibition of the ER-Golgi transport process with Brefeldin A leads to tumor regression. Our study collectively reveals the heterogeneity of PCa cell transcriptional response to androgen stimulation, demonstrates a functional role for increased ER-Golgi trafficking process, and provides a mechanism for how the process is augmented in PCa as well as the potential of targeting may provide novel treatment strategies.

2021-10-10 | GSE165562 | GEO

The interactome of the UapA transporter reveals putative new players in anterograde membrane cargo trafficking: UapA-DYDY TurboID

2023-11-21 | PXD043941 | Pride

The interactome of the UapA transporter reveals putative new players in anterograde membrane cargo trafficking: ΔartA: UapA-TurboID

2023-11-21 | PXD043944 | Pride

Expression analysis of pre-phenotype dt27J mouse dorsal root ganglia

2012-02-23 | GSE30960 | GEO