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

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 Po

Details

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

About the serial-volume-editor

Valérie Doye

Affiliations and Expertise

Institut Jacques Monod, Université Paris Diderot, France