Translation and Assembly of Radiolabeled Mitochondrial DNA-Encoded Protein Subunits from Cultured Cells and Isolated Mitochondria

Luke E Formosa, Annette Hofer, Christin Tischner, Tina Wenz, Michael T Ryan

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

2 Citations (Scopus)

Abstract

In higher eukaryotes, the mitochondrial electron transport chain consists of five multi-subunit membrane complexes responsible for the generation of cellular ATP. Of these, four complexes are under dual genetic control as they contain subunits encoded by both the mitochondrial and nuclear genomes, thereby adding another layer of complexity to the puzzle of respiratory complex biogenesis. These subunits must be synthesized and assembled in a coordinated manner in order to ensure correct biogenesis of different respiratory complexes. Here, we describe techniques to (1) specifically radiolabel proteins encoded by mtDNA to monitor the rate of synthesis using pulse labeling methods, and (2) analyze the stability, assembly, and turnover of subunits using pulse-chase methods in cultured cells and isolated mitochondria.
Original languageEnglish
Title of host publicationMitochondrial DNA
Subtitle of host publicationMethods and Protocols
EditorsMatthew McKenzie
Place of PublicationNew York NY USA
PublisherHumana Press
Pages115-130
Number of pages16
Edition3rd
ISBN (Electronic)978-1-4939-3040-1
ISBN (Print)978-1-4939-3039-5
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology: Springer Protocols
PublisherSpringer
Volume1351
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Cite this

Formosa, L. E., Hofer, A., Tischner, C., Wenz, T., & Ryan, M. T. (2016). Translation and Assembly of Radiolabeled Mitochondrial DNA-Encoded Protein Subunits from Cultured Cells and Isolated Mitochondria. In M. McKenzie (Ed.), Mitochondrial DNA: Methods and Protocols (3rd ed., pp. 115-130). (Methods in Molecular Biology: Springer Protocols; Vol. 1351). New York NY USA: Humana Press. https://doi.org/10.1007/978-1-4939-3040-1_9
Formosa, Luke E ; Hofer, Annette ; Tischner, Christin ; Wenz, Tina ; Ryan, Michael T. / Translation and Assembly of Radiolabeled Mitochondrial DNA-Encoded Protein Subunits from Cultured Cells and Isolated Mitochondria. Mitochondrial DNA: Methods and Protocols. editor / Matthew McKenzie. 3rd. ed. New York NY USA : Humana Press, 2016. pp. 115-130 (Methods in Molecular Biology: Springer Protocols).
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abstract = "In higher eukaryotes, the mitochondrial electron transport chain consists of five multi-subunit membrane complexes responsible for the generation of cellular ATP. Of these, four complexes are under dual genetic control as they contain subunits encoded by both the mitochondrial and nuclear genomes, thereby adding another layer of complexity to the puzzle of respiratory complex biogenesis. These subunits must be synthesized and assembled in a coordinated manner in order to ensure correct biogenesis of different respiratory complexes. Here, we describe techniques to (1) specifically radiolabel proteins encoded by mtDNA to monitor the rate of synthesis using pulse labeling methods, and (2) analyze the stability, assembly, and turnover of subunits using pulse-chase methods in cultured cells and isolated mitochondria.",
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Formosa, LE, Hofer, A, Tischner, C, Wenz, T & Ryan, MT 2016, Translation and Assembly of Radiolabeled Mitochondrial DNA-Encoded Protein Subunits from Cultured Cells and Isolated Mitochondria. in M McKenzie (ed.), Mitochondrial DNA: Methods and Protocols. 3rd edn, Methods in Molecular Biology: Springer Protocols, vol. 1351, Humana Press, New York NY USA, pp. 115-130. https://doi.org/10.1007/978-1-4939-3040-1_9

Translation and Assembly of Radiolabeled Mitochondrial DNA-Encoded Protein Subunits from Cultured Cells and Isolated Mitochondria. / Formosa, Luke E; Hofer, Annette; Tischner, Christin; Wenz, Tina; Ryan, Michael T.

Mitochondrial DNA: Methods and Protocols. ed. / Matthew McKenzie. 3rd. ed. New York NY USA : Humana Press, 2016. p. 115-130 (Methods in Molecular Biology: Springer Protocols; Vol. 1351).

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

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AU - Wenz, Tina

AU - Ryan, Michael T

PY - 2016

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N2 - In higher eukaryotes, the mitochondrial electron transport chain consists of five multi-subunit membrane complexes responsible for the generation of cellular ATP. Of these, four complexes are under dual genetic control as they contain subunits encoded by both the mitochondrial and nuclear genomes, thereby adding another layer of complexity to the puzzle of respiratory complex biogenesis. These subunits must be synthesized and assembled in a coordinated manner in order to ensure correct biogenesis of different respiratory complexes. Here, we describe techniques to (1) specifically radiolabel proteins encoded by mtDNA to monitor the rate of synthesis using pulse labeling methods, and (2) analyze the stability, assembly, and turnover of subunits using pulse-chase methods in cultured cells and isolated mitochondria.

AB - In higher eukaryotes, the mitochondrial electron transport chain consists of five multi-subunit membrane complexes responsible for the generation of cellular ATP. Of these, four complexes are under dual genetic control as they contain subunits encoded by both the mitochondrial and nuclear genomes, thereby adding another layer of complexity to the puzzle of respiratory complex biogenesis. These subunits must be synthesized and assembled in a coordinated manner in order to ensure correct biogenesis of different respiratory complexes. Here, we describe techniques to (1) specifically radiolabel proteins encoded by mtDNA to monitor the rate of synthesis using pulse labeling methods, and (2) analyze the stability, assembly, and turnover of subunits using pulse-chase methods in cultured cells and isolated mitochondria.

UR - http://www.ncbi.nlm.nih.gov/pubmed/26530678

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T3 - Methods in Molecular Biology: Springer Protocols

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Formosa LE, Hofer A, Tischner C, Wenz T, Ryan MT. Translation and Assembly of Radiolabeled Mitochondrial DNA-Encoded Protein Subunits from Cultured Cells and Isolated Mitochondria. In McKenzie M, editor, Mitochondrial DNA: Methods and Protocols. 3rd ed. New York NY USA: Humana Press. 2016. p. 115-130. (Methods in Molecular Biology: Springer Protocols). https://doi.org/10.1007/978-1-4939-3040-1_9