Main description:

This volume details the most recent advancements in the field of mitochondrial gene expression. Chapters guide readers through methods and protocols on mtDNA replication, transcription, and translation to membrane insertion of the mtDNA-encoded protein products. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Mitochondrial Gene Expression: Methods and Protocols aims to provide complementary approaches and practical guidelines.

Chapter 7 is available Open Access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Contents:

1. In vitro Analysis of mtDNA Replication

Jay P. Uhler and Maria Falkenberg

2. In vivo Analysis of mtDNA Replication at the Single Molecule Level and with High Resolution

Marco Tigano, Aaron Fraser Phillips, and Agnel Sfeir

3. In Vitro Reconstitution of Human Mitochondrial Transcription

Azadeh Sarfallah and Dmitry Temiakov

4. Investigating Mitochondrial Transcriptomes and RNA Processing using Circular RNA Sequencing

Irina Kuznetsova, Oliver Rackham, and Aleksandra Filipovska

5. Detection of 5-formylcytosine in Mitochondrial Transcriptome

Lindsey Van Haute and Michal Minczuk

6. Visualization of Mitochondrial RNA Granules in Cultured Cells using 5-Bromouridine Labeling

Vanessa Joanne Xavier and Jean-Claude Martinou

7. Quantitative Proteomics in Drosophila with Holidic Stable Isotope Labeling of Amino Acids in Fruit Flies (SILAF)

Florian A. Schober, Ilian Atanassov, Christoph Freyer, and Anna Wredenberg

8. Mass Spectrometric Analysis of Mitochondrial RNA Modifications

Yuma Ishigami, Tsutomu Suzuki, and Takeo Suzuki

9. mito- -Seq: A High-throughput Method for Systematic Mapping of Pseudouridine within Mitochondrial RNA

Aldema Sas-Chen, Ronit Nir, and Schraga Schwartz

10. High Throughput Detection of mtDNA Mutations Leading to tRNA Processing Errors

Marita Annika Isokallio and James Bruce Stewart

11. High-throughput Measurement of Mitochondrial RNA Turnover in Human Cultured Cells

Anna V. Kotrys, Lukasz S. Borowski, and Roman J. Szczesny

12. RNA Crosslinking to Analyze the Mitochondrial RNA-binding Proteome

S.L. van Esveld and J.N. Spelbrink

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

Matthew Zorkau, Yasmin Proctor-Kent,, Rolando Berlinguer-Palmini, Andrew Hamilton, Zofia M Chrzanowska-Lightowlers, and Robert N. Lightowlers

14. Mitoribosome Profiling From Human Cell Culture - A High Resolution View of Mitochondrial Translation

Pearce Sarah, Cipullo Miriam, Chung Betty, Brierley Ian, and Rorbach Joanna

15. Application of cryo-EM for Visualization of Mitoribosomes

Vivek Singh and A. Amunts

16. Sucrose Gradient Sedimentation Analysis of Mitochondrial Ribosomes

Austin Choi and Antoni Barrientos

17. The Analysis of Yeast Mitochondrial Translation

Andreas Carlstroem, Magdalena Rzepka, and Martin Ott

18. In situ Studies of Mitochondrial Translation by Cryoelectron Tomography

Robert Englmeier and Friedrich Foerster

19. Mitochondrial Complexome Profiling

Heiko Giese, Jana Meisterknecht, Juliana Heidler, and Ilka Wittig

20. Blue Native Electrophoresis to Study the Oxphos Complexes

Erika Fernandez-Vizarra and Massimo Zeviani

21. Identification of Putative Mitochondrial Protease Substrates

Eduard Hofsetz, Pitter F. Huesgen, and Aleksandra Trifunovic