Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Complexome profiling of mitochondrial protein complexes from CRISPR/Cas9 edited U2OS cells lacking COA5

ABSTRACT: Mitochondria are complex cellular organelles that harness the process of oxidative phosphorylation (OXPHOS) to provide a usable energy source for the cell and body. The failure of mitochondria to achieve this is associated with a wide spectrum of disorders that affect both children and adults with an incidence of at least 1/4300 1. Due to the dual genetic control of genes encoding the structural components of the mitochondrial OXPHOS system, mitochondrial disorders can occur via almost any pattern of inheritance. The assembly of mitochondrial OXPHOS complexes is an intricate process and requires tight co-regulation of two evolutionarily distinct gene expression machineries. In addition, early- and late-stage ancillary chaperones have essential roles in coordinating OXPHOS assembly, regulating the step-wise insertion of individual subunits and /or essential co-factors. Mitochondrial OXPHOS complex IV (cytochrome c oxidase) comprises of 14 subunits, with all three catalytic subunits (COX1, COX2, and COX3) encoded by mitochondrial DNA. Complex IV represents the final stage of electron transfer across the electron transport chain by cytochrome c to oxygen as the final electron acceptor in OXPHOS. Previous work in our lab and others identified pathogenic variants in a novel complex IV assembly factor COA5 associated with cardiomyopathy and defective complex IV assembly. Here we used complexome profiling to study the role of COA5 in early stage complex IV assembly using a CRISPR/Cas9-mediated COA5 gene knockout cell line.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Ilka Wittig

LAB HEAD: Robert Taylor

PROVIDER: PXD053461 | Pride | 2025-01-07

REPOSITORIES: pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	BN_labeled.tif	Other
	Data_analysis.xlsx	Xlsx
	Gel.jpg	Other
	P22_075_Taylor_Ctrl_01.raw	Raw
	P22_075_Taylor_Ctrl_02.raw	Raw

Items per page:

1 - 5 of 101

Similar Datasets

Complexome profiling of mitochondrial protein complexes in single and double MIC26 and MIC27 knockout cells

Project description:Here we analysed the impact of two subunits of the mitochondrial contact site and cristae organizing system (MICOS) complex, MIC26 and MIC27, on the assembly and stability of mitochondrial complexes of the oxidative phosphorylation system (OXPHOS).

2020-07-28 | PXD016733 | Pride

Complexome profiling of human cybrids laking mitochondrial genes Cox1 or Cox2

Project description:Mitochondrial respiratory chain (MRC) complexes I, III and IV are associated into large molecular structures named supercomplexes (SCs) or respirasomes, whose functional roles remain to be fully understood. Biochemical analyses in a diversity of OXPHOS-deficient cellular and animal models support the idea that one of the SCs function is to confer stability to individual MRC complexes, in particular to complex I (CI). To gain insight into the mechanisms that regulate the structural interdependences among MRC complexes, we performed complexome profiling of human cybrids laking mitochondrial genes Cox1 or Cox2 and compared protein assemblies and intermediates to control 143B cells.

2020-07-01 | PXD017840 | Pride

Complexome profiling of MICOS complex, impact of subunits MIC26 and MIC27

Project description:Here we analysed the impact of two subunits (MIC26 and MIC27) on the assembly and stability of the mitochondrial contact site and cristae organizing system (MICOS) complex.

2020-07-28 | PXD016732 | Pride

Native Prohibitin Complex Enrichment

Project description:Folding of newly synthesized proteins poses challenges for a functional proteome. Dedicated protein quality control (PQC) systems promote either the folding of nascent polypeptides at ribosomes or, if this fails, ensure their degradation. While well studied for cytosolic protein biogenesis, it is not understood how these processes work for mitochondrially-encoded proteins, key subunits of the oxidative phosphorylation system (OXPHOS). Here, we identify dedicated hubs in proximity to mitoribosomal tunnel exits coordinating mitochondrial translation, protein import, assembly, and protein turnover. Conserved prohibitin/m-AAA protease supercomplexes and the availability of assembly chaperones determine the fate of newly synthesized proteins by molecular triaging. The localization of these competing activities in vicinity to the mitoribosomal tunnel exit allows for a prompt decision whether newly synthesized proteins are fed into OXPHOS assembly or degraded. This repository contains the results of the FLAG-based enrichment of the native prohibitin complex. Please note that we measured elution either with LDS sample buffer or using FLAG-peptides. In the published study, the LDS elution buffer was shown.

2023-08-14 | PXD043671 | Pride

A stop-and-go mechanism controls mitochondrial respiration

Project description:Oxidative phosphorylation (OXPHOS) is essential for the synthesis of the vast majority of ATP in eukaryotic cells. It is carried out by multi-protein assemblies that form the electron transport chain (ETC), which are coupled to the mitochondrial ATP synthase, which uses the proton gradient generated by the ETC to drive ATP synthesis. The assembly of the OXPHOS protein machinery requires the coordinated integration of proteins encoded in the nuclear and the mitochondrial genomes, imposing an additional layer of complexity. While the biogenesis of the individual OXPHOS complexes is well understood, it remains unknown how the assembly of the ETC and ATP synthase is coordinated to achieve the correct stoichiometry of the OXPHOS machinery. Here, we identify the mitochondrial regulatory hub for respiratory assembly (MiRA), which serves as a platform on which complex IV and complex V biogenesis are synchronised to ensure balanced assembly of the ETC and complex V. At the molecular level, this is achieved by a stop-and-go safeguarding mechanism: The novel mitochondrial protein Mra1 binds to and slows down complex IV assembly. Two Go signals are then required for assembly to proceed: Binding of the complex IV assembly factor Rcf2 and interaction with the mitoribosome, which translates the complex V subunit Atp9. Both Go signals induce the clipping and subsequent degradation of Mra1, relieving the molecular break and allowing parallel maturation of complexes IV and V. The absence of the stop-and-go safety mechanism results in the formation of immature complexes, decreased ATP levels and reduced cell viability. The antagonistic activities at MiRA provide a regulated orchestration of complex IV and V assembly which is essential to avoid the predominance of either complex and an unbalanced ratio of OXPHOS complexes, thus ensuring the correct stoichiometry of protein machineries encoded by two different genomes.

2024-02-16 | PXD044800 | Pride

Defective mitochondrial COX1 translation due to loss of COX14 function triggers ROS-induced inflammation in mouse liver

Project description:Mitochondrial oxidative phosphorylation (OXPHOS) fuels cellular ATP demands. OXPHOS defects lead to severe human disorders with unexplained tissue specific pathologies. Mitochondrial gene expression is essential for OXPHOS biogenesis since core subunits of the complexes are mitochondrial-encoded. COX14 is required for translation of COX1, the central mitochondrial-encoded subunit of complex IV. Here we describe a COX14 mutant mouse corresponding to a patient with complex IV deficiency. COX14M19I mice display broad tissue-specific pathologies. A hallmark phenotype is severe liver inflammation linked to release of mitochondrial RNA into the cytosol sensed by RIG-1 pathway. We find that mitochondrial RNA release is triggered by increased reactive oxygen species production in the deficiency of complex IV. Additionally, we describe a COA3Y72C mouse, affected in an assembly factor that cooperates with COX14 in early COX1 biogenesis, which displays a similar yet milder inflammatory phenotype. Our study provides insight into a link between defective mitochondrial gene expression and tissue-specific inflammation.

2024-06-11 | GSE269535 | GEO

Complexome profiling of mitochondrial protein complexes from patients with mutation in NDUFC2 gene

Project description:NDUFC2 is a subunit of the membrane part of NADH:ubiquinone oxidoreductase (complex I). Here we used complexome profiling to study the role of NDUFC2 in complex I stability and assembly.

2020-08-10 | PXD014936 | Pride

Complexome profiling of brain mitochondrial membranes from Clpp-/- mouse

Project description:The autosomal recessive Perrault syndrome with juvenile ovarian failure, progressive sensorineural deafness, ataxia and leukoencephalopathy can be caused by loss of function mutations in CLPP. This gene encodes a peptidase that is conserved since bacteria and localizes to mitochondrial matrix in eukaryotes. Here, assembly and stability of mitochondrial complexes from adult Clpp-/- mouse brains were analyzed by Complexome profiling.

2022-02-17 | PXD025478 | Pride

Complexome profiling of BRAWNIN KO mouse heart mitochondria

Project description:BRAWNIN (BR) is a mitochondrial microprotein important for assembly of complex III. Here we used complexome to study the composition of supercomplexes in BR KO mouse heart mitochondria. BR deficiency induces the formation of a higher order electron transport chain complex which we renamed as extra-large supercomplex (SC-XL). The stoichiometry of SC-XL is I2+III2. SC-XL formation in the BR KO system alleviates the CIII defects and promotes compensatory OXPHOS increase. It is of higher electron transport efficiency and of lower ROS generation.

2025-01-08 | PXD044957 | Pride

Regulators of mitonuclear balance link mitochondrial metabolism to mtDNA expression

Project description:Mitochondrial oxidative phosphorylation (OXPHOS) complexes are assembled from proteins encoded by both nuclear and mitochondrial DNA. These dual-origin enzymes pose a complex gene regulatory challenge for cells requiring coordinated gene expression across organelles. To identify genes involved in dual-origin protein complex synthesis, we performed FACS-based genome-wide screens analyzing mutant cells with unbalanced levels of mitochondrial- and nuclear-encoded subunits of Complex IV. We identified novel genes involved in OXPHOS biogenesis, including two uncharacterized genes: PREPL and NME6. We found that PREPL specifically impacts Complex IV biogenesis by acting at the intersection of mitochondrial lipid metabolism and protein synthesis, while NME6, an uncharacterized nucleoside diphosphate kinase (NDPK), controls OXPHOS biogenesis through multiple mechanisms reliant on its NDPK domain. First, NME6 forms a complex with RCC1L, which together perform NDPK activity to maintain local mitochondrial pyrimidine triphosphate levels essential for mitochondrial RNA abundance. Second, NME6 modulates the activity of mitoribosome regulatory complexes, altering mitoribosome assembly and mitochondrial RNA pseudouridylation. Taken together, we propose that NME6 acts as a link between compartmentalized mitochondrial metabolites and mitochondrial gene expression.

2023-08-17 | GSE240576 | GEO