Proteome and cytoskeleton responses in osteosarcoma cells with reduced OXPHOS activity

Annunen-Rasila, Johanna; Ohlmeier, Steffen; Tuokko, Hanna; Veijola, Johanna; Majamaa, Kari

doi:10.1002/pmic.200601031

Cited by 27 publications

(25 citation statements)

References 59 publications

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“…It has already been reported half a century ago, that mitochondrial morphology is dependent on the respiration status (57). Furthermore, immunolabeling studies revealed that 0 cells have severe cytoskeletal alterations, such as a collapsed vimentin network, a dense accumulation of mitochondria around the nucleus and only a few mitochondria distributed throughout the cytoplasm (58). In 0 cells, the reticulum appears disrupted, that yields in a distribution of small individual organelles instead of reticular networks with tubular cristae (59 -61).…”

Section: Discussionmentioning

confidence: 93%

An Impaired Respiratory Electron Chain Triggers Down-regulation of the Energy Metabolism and De-ubiquitination of Solute Carrier Amino Acid Transporters

Aretz

Hardt

Wittig

et al. 2016

Molecular & Cellular Proteomics

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Section: Discussionmentioning

confidence: 93%

An Impaired Respiratory Electron Chain Triggers Down-regulation of the Energy Metabolism and De-ubiquitination of Solute Carrier Amino Acid Transporters

Aretz

Hardt

Wittig

et al. 2016

Molecular & Cellular Proteomics

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“…One argument is that in various cell lines, intermediate filaments are enriched in the mitochondrial fraction and coimmunoprecipitate with intact mitochondria (11). In addition, a disturbed intermediate filament network was found in myoblasts from the patients with the heteroplasmic m.3243A→G mutation (12), and in cultured skin fibroblasts from the patients with the m.5650G→A mutation in the mitochondrial tRNA Ala gene (13). In these studies, the distribution of actin and α-tubulin was unchanged.…”

Section: Articlesmentioning

confidence: 88%

Fluorescence imaging of mitochondria in cultured skin fibroblasts: a useful method for the detection of oxidative phosphorylation defects

et al. 2012

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Background: Protons are pumped from the mitochondrial matrix via oxidative phosphorylation (OXPhOs) into the intermembrane space, creating an electric membrane potential (ΔΨ) that is used for adenosine triphosphate (aTP) production. Defects in one or more of the OXPhOs complexes are associated with a variety of clinical symptoms, often making it difficult to pinpoint the causal mutation. Methods: In this article, a microscopic method for the quantitative evaluation of ΔΨ in cultured skin fibroblasts is described. The method using 5,5′,6,6′-tetraethylbenzimidazolyl-carbocyanine iodide (Jc-1) fluorescence staining was tested in a selection of OXPhOs-deficient cell lines. results: a significant reduction of ΔΨ was found in the cell lines of patients with either an isolated defect in complex I, II, or IV or a combined defect (complex I + complex IV). ΔΨ was not reduced in the fibroblasts of two patients with severe complex V deficiency. addition of the complex I inhibitor rotenone induced a significant reduction of ΔΨ and perinuclear relocalization of the mitochondria. In cells with a heteroplasmic mitochondrial DNa (mtDNa) defect, a more heterogeneous reduction of ΔΨ was detected. conclusion: Our data show that imaging of ΔΨ in cultured skin fibroblasts is a useful method for the evaluation of OXPhOs functioning in cultured cell lines.t he majority of cellular adenosine triphosphate (ATP) is supplied in the mitochondria through oxidative phosphorylation (OXPHOS). The OXPHOS system consists of five multiprotein complexes embedded within the inner mitochondrial membrane: complex I (reduced nicotinamide adenine dinucleotide: ubiquinone oxidoreductase), complex II (succinate: ubiquinone oxidoreductase), complex III (ubiquinol: cytochrome c oxidoreductase), complex IV (cytochrome c oxidase), and complex V (ATP synthase). Complexes I, III, and IV pump protons, donated by reduced nicotinamide adenine dinucleotide, ubiquinone, and cytochrome c, respectively, into the mitochondrial intermembrane space. The proton gradient that develops between the mitochondrial matrix and the intermembrane space generates an electrochemical membrane potential (ΔΨ), which is the driving force for the conversion of ADP and inorganic phosphate into ATP (1).The estimated incidence of OXPHOS defects in humans is ~1 in 5,000 live births. Isolated complex I deficiency (2) and the mitochondrial DNA (mtDNA) depletion syndromes (3) are the most commonly recognized mitochondrial disorders. Defects in OXPHOS are associated with a broad spectrum of symptoms and syndromes, ranging from mild myopathy to severe multisystem disorders. OXPHOS defects are complex due to the dual origin of genes involved and the specific nature of the mitochondrial genome. The mtDNA encodes 13 structural proteins of the complexes I, III, IV, and V; two rRNAs; and 22 transfer RNAs (tRNAs) necessary for intramitochondrial protein translation. The mitochondrial genome is polyploid, with multiple copies of mtDNA present within each individual mitochondrion. Two different populatio...

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“…A third class of cytoskeletal structures is the intermediate filaments, typified by the protein vimentin (94). Vimentin has been implicated in diverse functions, from structural roles to roles in the cell cycle and mitosis (94), as well as lipid metabolism, particularly in regard to the biology of lipid droplets (76), whose roles in lipid storage and metabolism are the likely link to energy metabolism and mitochondrial function (3). More relevant to this study, vimentin is reorganized around the PV in T. gondii-infected cells (15,38).…”

Section: Vol 76 2008 Modulation Of the Host Cell Proteome By T Gonmentioning

confidence: 99%

Modulation of the Host Cell Proteome by the Intracellular Apicomplexan ParasiteToxoplasma gondii

et al. 2008

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To investigate how intracellular parasites manipulate their host cell environment at the molecular level, we undertook a quantitative proteomic study of cells following infection with the apicomplexan parasite Toxoplasma gondii. Using conventional two-dimensional electrophoresis, difference gel electrophoresis (DIGE), and mass spectrometry, we identified host proteins that were consistently modulated in expression following infection. We detected modification of protein expression in key metabolic pathways, including glycolysis, lipid and sterol metabolism, mitosis, apoptosis, and structural-protein expression, suggestive of global reprogramming of cell metabolism by the parasite. Many of the differentially expressed proteins had not been previously implicated in the response to the parasite, while others provide important corroborative protein evidence for previously proposed hypotheses of pathogen-cell interactions. Significantly, over one-third of all modulated proteins were mitochondrial, and this was further investigated by DIGE analysis of a mitochondrion-enriched preparation from infected cells. Comparison of our proteomic data with previous transcriptional studies suggested that a complex relationship exits between transcription and protein expression that may be partly explained by posttranslational modifications of proteins and revealed the importance of investigating protein changes when interpreting transcriptional data. To investigate this further, we used phosphatase treatment and DIGE to demonstrate changes in the phosphorylation states of several key proteins following infection. Overall, our findings indicate that the host cell proteome responds in a dramatic way to T. gondii invasion, in terms of both protein expression changes and protein modifications, and reveal a complex and intimate molecular relationship between host and parasite.The obligate intracellular protozoan Toxoplasma gondii has evolved an intimate relationship with its host that extends to the cellular and molecular levels (82, 83). The absolute requirement for invasion of an appropriate host cell in the replication of the parasite suggests that modification of host functions is central to pathogenesis. Intuitively, this subversion of the cell must be a complex process, since host cells are not inherently programmed to provide an environment conducive to pathogens. Host cells have evolved primary lines of defense as countermeasures to pathogen invasion, establishment, and replication. These include elaborate systems, such as the phagolysosomal fusion, reactive oxygen and nitrogen intermediates, the sequestration of nutrients, and even cell suicide (apoptosis), as defenses to limit pathogen growth. The implementation of these systems and the ability of successful pathogens to mitigate their effects are ultimately mediated by changes in both the levels and activities of key proteins. While some of these changes may be transcriptionally regulated and thus potentially revealed by microarray analysis, the true effectors are proteins,...

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Proteome and cytoskeleton responses in osteosarcoma cells with reduced OXPHOS activity

Cited by 27 publications

References 59 publications

An Impaired Respiratory Electron Chain Triggers Down-regulation of the Energy Metabolism and De-ubiquitination of Solute Carrier Amino Acid Transporters

An Impaired Respiratory Electron Chain Triggers Down-regulation of the Energy Metabolism and De-ubiquitination of Solute Carrier Amino Acid Transporters

Fluorescence imaging of mitochondria in cultured skin fibroblasts: a useful method for the detection of oxidative phosphorylation defects

Modulation of the Host Cell Proteome by the Intracellular Apicomplexan ParasiteToxoplasma gondii

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