The ESCRT Complexes, 1st ed. 2019
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1998

Language: English

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The ESCRT Complexes
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The ESCRT Complexes
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322 p. · 17.8x25.4 cm · Hardback
This detailed collection gathers both established and recent technical procedures to study the Endosomal Sorting Complex Required for Transport (ESCRT) complexes in a wide range of biological systems: Archaea, A. thaliana, U. maydis, S. cerevisiae, S. pombe, C. elegans, D. melanogaster, and mammalian cells. Opening with a section on imaging techniques, the book continues with chapters covering biochemical approaches presenting strategies for production and characterization of recombinant ESCRT proteins, or of specific ESCRT protein domains, as well as genetic and proteomic experimental approaches. Written for the highly successful Methods in Molecular Biology series, chapters include introduction 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 practical, The ESCRT Complexes: Methods and Protocols serves as a compact guide for researchers interested in establishing an integrated approach to investigate the ESCRT machinery functions in cell biology.

1. Analysis of the Archaeal ESCRT Apparatus

            Rachel Y. Samson, Iain G. Duggin, and Stephen D. Bell

 

2. Genetic and Cell Biology Methods to Study ESCRTs in Drosophila melanogaster

            Marco Gualtieri and Thomas Vaccari

 

3. Functional Analysis of ESCRT-Positive Extracellular Vesicles in the Drosophila Wing Imaginal Disc

            Tamas Matusek, Pascal Thérond, and Maximilian Fürthauer

 

4. Subcellular Localization of ESCRT-II in the Nematode C. elegans by Correlative-Light Electron Microscopy

            Céline Largeau, Emmanuel Culetto, and Renaud Legouis

 

5. Proximity Ligation Assay (PLA) to Determine the Endosomal Localization of ESCRT Subunit in Virus-Infected Cells

            Binod Kumar, Mohanan Valiya Veettil, Arunava Roy, and Bala Chandran

 

6. Immuno-Localization of ESCRT Proteins in Virus-Infected Cells by Fluorescence and Electron Microscopy

            Keisuke Tabata, Atsuki Nara, Hiroko Omori, and Eiji Morita

 

7. Single Cell Fluorescence Ratio Image Analysis for Studying ESCRT Function in Receptor Trafficking

            Jalal M. Kazan, Gergely L. Lukacs, Pirjo M. Apaja, and Arnim Pause

 

8. Genetic and Biochemical Analyses of Yeast ESCRT

            Sudeep Banjade, Shaogeng Tang, and Scott D. Emr

 

9. Live Imaging of ESCRT Proteins in Microfluidically-Isolated Hippocampal Axons

            Veronica Birdsall, Jose C. Martinez, Lisa Randolph, Ulrich Hengst, and Clarissa L. Waites

 

10. Studying the Spatial Organization of ESCRTs in Cytokinetic Abscission Using the High Resolution Imaging Techniques SIM and Cryo-SXT

            Shai Adar-Levor, Inna Goliand, Michael Elbaum, and Natalie Elia

 

11. Three Dimensional Surface Rendering of ESCRT Proteins Microscopy Data Using UCSF Chimera Software

            Romain Le Bars, Michele W. Bianchi, and Christophe Lefebvre

 

12. Transient Expression of ESCRT-Components in Arabidopsis Root Cell Suspension Culture-Derived Protoplasts

            Marie-Kristin Nagel, Karin Vogel, and Erika Isono

 

13. Crystallization and Biophysical Approaches for Studying the Interactions between the Vps4-MIT Domain and ESCRT-III Proteins

            Takayuki Obita, Rieko Kojima, and Mineyuki Mizuguchi

 

14. Biochemical Approaches to Studying C. elegans ESCRT Function In Vitro

            Samuel Block, Amber L. Schuh, and Anjon Audhya

 

15. Purification of Recombinant ESCRT-III Proteins and Their Use in Atomic Force Microscopy and In Vitro Binding and Phosphorylation Assays

            Luisa Capalbo, Ioanna Mela, Maria Alba Abad, A. Arockia Jeyaprakash, J. Michael Edwardson, and Pier Paolo D’Avino

 

16. Assessment of ESCRT Protein CHMP5 Activity on Client Protein Ubiquitination by Immunoprecipitation and Western Blotting

            Francheska Son, Katharine Umphred-Wilson, Jae-Hyuck Shim, and Stanley Adoro

 

17. Purification of Plant ESCRT Proteins for Polyclonal Antibody Production

            Julio Paez-Valencia and Marisa S. Otegui

 

18. Genetic and Cytological Methods to Study ESCRT Cell Cycle Function in Fission Yeast

            Imane M. Rezig, Shaun K. Bremner, Musab S. Bhutta, Ian P. Salt, Gwyn W. Gould, and Christopher J. McInerny

 

19. ESCRT Mutant Analysis and Imaging of ESCRT Components in the Model Fungus Ustilago maydis

            Carl Haag, Thomas Klein, and Michael Feldbrügge

 

20. Genetic Suppressor Screen Using an Inducible FREE1-RNAi Line to Detect ESCRT Genetic Interactors in Arabidopsisthaliana

            Qiong Zhao, Ying Zhu, Wenhan Cao, Jinbo Shen, Yong Cui, Shuxian Huang, and Liwen Jiang

 

21. Screening of Interactions with the ESCRT Machinery by a Gaussia princeps Split Luciferase-Based Complementation Assay

            Rina Barouch-Bentov, Yves Jacob, and Shirit Einav

 

22. RNA Interference-Mediated Inhibition of ESCRT in Mammalian Cells

            Katherine Bowers

Includes cutting-edge techniques

Provides step-by-step detail essential for reproducible results

Contains key implementation advice from the experts