Proteomics

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Identification of proteins differentially regulated upon perturbed mitochondrial gene expression


ABSTRACT: The aim of this project was to search for the new factors engaged in mitochondrial nucleic acids metabolism under stress conditions. To this end we treated human 293 cells with mitochondrial replication/transcription inhibitor ethidium bromide and we compared proteomes of untreated vs treated cells. We were looking for the proteins whose level would be differentially regulated in response to disturbed mitochondrial gene expression. In our studies we utilized a stable 293 cell line that inducible expresses a fusion of EGFP with mitochondria targeting sequence. This approach enabled us to control mitochondrial import by analysis of the abundance of mitochondrial EGFP marker. We isolated highly purified mitochondria from the cells, lysed them and subjected protein extracts to labelling with iTRAQ reagents. We performed quantitative analysis of stress- induced changes of human mitochondrial proteome with the use of MaxQuant software.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Dominik Cysewski  

LAB HEAD: Roman J. Szczesny

PROVIDER: PXD012055 | Pride | 2019-06-13

REPOSITORIES: Pride

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Action DRS
70715cys_A5h.raw Raw
70715cys_Aa.raw Raw
70715cys_Ab.raw Raw
70715cys_B5h.raw Raw
70715cys_Ba.raw Raw
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Publications

Quantitative proteomics revealed C6orf203/MTRES1 as a factor preventing stress-induced transcription deficiency in human mitochondria.

Kotrys Anna V AV   Cysewski Dominik D   Czarnomska Sylwia D SD   Pietras Zbigniew Z   Borowski Lukasz S LS   Dziembowski Andrzej A   Szczesny Roman J RJ  

Nucleic acids research 20190801 14


Maintenance of mitochondrial gene expression is crucial for cellular homeostasis. Stress conditions may lead to a temporary reduction of mitochondrial genome copy number, raising the risk of insufficient expression of mitochondrial encoded genes. Little is known how compensatory mechanisms operate to maintain proper mitochondrial transcripts levels upon disturbed transcription and which proteins are involved in them. Here we performed a quantitative proteomic screen to search for proteins that s  ...[more]

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