Nuclear Pore Complexes and Nucleocytoplasmic Transport - Methods - 1st Edition - ISBN: 9780124171602, 9780124171787

Nuclear Pore Complexes and Nucleocytoplasmic Transport - Methods, Volume 122

1st Edition

Serial Volume Editors: Valérie Doye
eBook ISBN: 9780124171787
Hardcover ISBN: 9780124171602
Imprint: Academic Press
Published Date: 15th May 2014
Page Count: 554
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Table of Contents

  • Preface
  • Chapter 1: Fifty Years of Nuclear Pores and Nucleocytoplasmic Transport Studies: Multiple Tools Revealing Complex Rules
    • Abstract
    • Introduction
    • Concluding Remarks
    • Acknowledgments
  • Chapter 2: Imaging Metazoan Nuclear Pore Complexes by Field Emission Scanning Electron Microscopy
    • Abstract
    • Introduction
    • Acknowledgments
  • Chapter 3: Imaging Yeast NPCs: From Classical Electron Microscopy to Immuno-SEM
    • Abstract
    • Introduction
    • Conclusions and Perspectives
    • Acknowledgments
  • Chapter 4: Exploring Nuclear Pore Complex Molecular Architecture by Immuno-Electron Microscopy Using Xenopus Oocytes
    • Abstract
    • Introduction
    • 4.1 Materials
    • 4.2 Experimental Strategies (Fig. 4.2)
    • 4.3 Preparation of Antibodies Conjugated with Colloidal Gold Particles
    • 4.4 Immunogold Labeling of Nucleoporins Using Anti-Nucleoporin Antibodies
    • 4.5 Immunogold Labeling of Nucleoporins Using Epitope-Tagged Nucleoporins
    • Concluding Remarks
    • Acknowledgments
  • Chapter 5: Utilizing the Dyn2 Dimerization-Zipper as a Tool to Probe NPC Structure and Function
    • Abstract
    • Introduction
    • Conclusion
    • Acknowledgment
  • Chapter 6: The Use of Targeted Proteomics to Determine the Stoichiometry of Large Macromolecular Assemblies
    • Abstract
    • Introduction
    • Conclusions
    • Acknowledgments
  • Chapter 7: A Pulse–Chase Epitope Labeling to Study Cellular Dynamics of Newly Synthesized Proteins: A Novel Strategy to Characterize NPC Biogenesis and Ribosome Maturation/Export
    • Abstract
    • Introduction
    • Conclusion and Perspectives
    • Acknowledgments
  • Chapter 8: Analysis of Nuclear Reconstitution, Nuclear Envelope Assembly, and Nuclear Pore Assembly Using Xenopus In Vitro Assays
    • Abstract
    • Introduction
    • 8.3 Demembranated Sperm Chromatin
    • 8.4 In vitro Reconstitution of Nuclei
    • 8.5 Assaying Assembly and Integrity of the Nuclear Envelope
    • 8.6 A Nuclear Pore Complex Assembly Assay Using Pore-Free Nuclear Intermediates
    • Conclusion
  • Chapter 9: Xenopus In Vitro Assays to Analyze the Function of Transmembrane Nucleoporins and Targeting of Inner Nuclear Membrane Proteins
    • Abstract
    • Introduction
    • Conclusion
  • Chapter 10: Imaging the Assembly, Structure, and Function of the Nuclear Pore Inside Cells
    • Abstract
    • Introduction
    • Acknowledgments
  • Chapter 11: Cell-Fusion Method to Visualize Interphase Nuclear Pore Formation
    • Abstract
    • Introduction
    • Conclusions
    • Acknowledgments
  • Chapter 12: An In Vitro System to Study Nuclear Envelope Breakdown
    • Abstract
    • Introduction
    • Acknowledgments
  • Chapter 13: Modern Tools to Study Nuclear Pore Complexes and Nucleocytoplasmic Transport in Caenorhabditis elegans
    • Abstract
    • Introduction
    • 13.1 Forward and Reverse Genetics
    • 13.2 Transgenesis
    • 13.3 Live Imaging of Embryos
    • 13.4 In vivo methods to evaluate structural and functional integrity of the NE
    • 13.5 Immunofluorescence and Electron Microscopy
    • 13.6 Interaction of Nups with Chromatin
    • Summary and Future Perspectives
    • Acknowledgments
  • Chapter 14: Assessing Regulated Nuclear Transport in Saccharomyces cerevisiae
    • Abstract
    • Introduction
    • Acknowledgments
  • Chapter 15: Analysis of Nucleocytoplasmic Transport in Digitonin-Permeabilized Cells Under Different Cellular Conditions
    • Abstract
    • Introduction
    • Conclusions and Perspectives
    • Acknowledgments
  • Chapter 16: Novel Approaches for the Identification of Nuclear Transport Receptor Substrates
    • Abstract
    • Introduction
    • Conclusions
    • Acknowledgments
  • Chapter 17: NPC Mimics: Probing the Mechanism of Nucleocytoplasmic Transport
    • Abstract
    • Introduction
    • Conclusions
  • Chapter 18: Analysis of RNA Transport in Xenopus Oocytes and Mammalian Cells
    • Abstract
    • Introduction
    • Concluding Remarks
    • Acknowledgment
  • Chapter 19: Strategies for Investigating Nuclear–Cytoplasmic tRNA Dynamics in Yeast and Mammalian Cells
    • Abstract
    • Introduction
    • Concluding remarks
  • Chapter 20: Dissecting Ribosome Assembly and Transport in Budding Yeast
    • Abstract
    • Introduction
    • Conclusions
    • Acknowledgments
  • Chapter 21: Approaches to Studying Subnuclear Organization and Gene–Nuclear Pore Interactions
    • Abstract
    • Introduction
    • Concluding Remarks, Possible Caveats, and Troubleshooting
    • Acknowledgments
  • Index
  • Volumes in Series

Description

Volume 122 of Methods in Cell Biology describes modern tools and techniques used to study nuclear pore complexes and nucleocytoplasmic transport in diverse eukaryotic model systems (including mammalian cells, Xenopus, C. elegans, yeast).

The volume enables investigators to analyze nuclear pore complex structure, assembly, and dynamics; to evaluate protein and RNA trafficking through the nuclear envelope; and to design in vivo or in vitro assays appropriate to their research needs.

Beyond the study of nuclear pores and transport as such, these protocols will also be helpful to scientists characterizing gene regulation, signal transduction, cell cycle, viral infections, or aging. The NPC being one of the largest multiprotein complexes in the cell, some protocols will also be of interest for people currently characterizing other macromolecular assemblies. This book is thus designed for laboratory use by graduate students, technicians, and researchers in many molecular and cellular disciplines.

Key Features

  • Describes modern tools and techniques used to study nuclear pore complexes and nucleocytoplasmic transport in diverse eukaryotic model systems (mammalian cells, Xenopus, C. elegans, yeast)
  • Chapters are written by experts in the field
  • Cutting-edge material

Readership

Researchers and students in cell, molecular and developmental biology


Details

No. of pages:
554
Language:
English
Copyright:
© Academic Press 2014
Published:
Imprint:
Academic Press
eBook ISBN:
9780124171787
Hardcover ISBN:
9780124171602

Ratings and Reviews


About the Serial Volume Editors

Valérie Doye Serial Volume Editor

Affiliations and Expertise

Institut Jacques Monod, Université Paris Diderot, France