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Visualizing Mitochondrial Ribosomal RNA and Mitochondrial Protein Synthesis in Human Cell Lines

Lookup NU author(s): Matt Zorkau, Yasmin Proctor-Kent, Dr Rolando Berlinguer Palmini, Professor Zofia Chrzanowska-LightowlersORCiD, Professor Robert Lightowlers


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© 2021, Springer Science+Business Media, LLC, part of Springer Nature.Human mitochondria contain their own DNA (mtDNA) that encodes 13 proteins all of which are core subunits of oxidative phosphorylation (OXPHOS) complexes. To form functional complexes, these 13 components need to be correctly assembled with approximately 70 nuclear-encoded subunits that are imported following synthesis in the cytosol. How this complicated coordinated translation and assembly is choreographed is still not clear. Methods are being developed to determine whether all members of a particular complex are translated in close proximity, whether protein synthesis is clustered in submitochondrial factories, whether these align with incoming polypeptides, and if there is evidence for co-translational translation that is regulated and limited by the interaction of the incoming proteins with synthesis of their mtDNA-encoded partners. Two methods are described in this chapter to visualize the distribution of mitochondrial ribosomal RNAs in conjunction with newly synthesized mitochondrial proteins. The first combines RNA Fluorescent In Situ Hybridization (FISH) and super-resolution immunocytochemistry to pinpoint mitochondrial ribosomal RNA. The second localizes nascent translation within the mitochondrial network through non-canonical amino acid labeling, click chemistry and fluorescent microscopy.

Publication metadata

Author(s): Zorkau M, Proctor-Kent Y, Berlinguer-Palmini R, Hamilton A, Chrzanowska-Lightowlers ZM, Lightowlers RN

Publication type: Book Chapter

Publication status: Published

Book Title: Methods in Molecular Biology

Year: 2021

Volume: 2192

Pages: 159-181

Online publication date: 24/11/2020

Acceptance date: 02/04/2016

Publisher: Humana Press Inc.


DOI: 10.1007/978-1-0716-0834-0_13

PubMed id: 33230773

Library holdings: Search Newcastle University Library for this item

ISBN: 9781071608340