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.
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Table of Contents
1. Fifty Years of Nuclear Pores and Nucleocytoplasmic Transport Studies: Multiple Tools Revealing Complex Rules 2. Imaging Metazoan Nuclear Pore Complexes by Field Emission Scanning Electron Microscopy 3. Imaging Yeast NPCs: From Classical Electron Microscopy to Immuno SEM 4. Exploring Nuclear Pore Complex Molecular Architecture by Immuno-Electron Microscopy using Xenopus Oocytes 5. Utilizing the Dyn2 Dimerization-Zipper as a Tool to Probe NPC Structure and Function 6. The use of Targeted Proteomics to Determine the Stoichiometry of Large Macromolecular Assemblies 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 8. Analysis of Nuclear Reconstitution, Nuclear Envelope Assembly, and Nuclear Pore Assembly using Xenopus in vitro Assays 9. Xenopus in vitro Assays to Analyze the Function of Transmembrane Nucleoporins and Targeting of Inner Nuclear Membrane Proteins 10. Imaging the Assembly, Structure and Function of the Nuclear Pore Inside Cells 11. Cell-Fusion Method to Visualize Interphase Nuclear Pore (NPC) Formation 12. An in vitro System to Study Nuclear Envelope Breakdown 13. Modern Tools to Study Nuclear Pore Complexes and Nucleocytoplasmic Transport in Caenorhabditis elegans 14. Assessing Regulated Nuclear Transport in Saccharomyces cerevisiae 15. Analysis of Nucleocytoplasmic Transport in Digitonin-Permeabilized Cells under Different Cellular Conditions 16. Novel Approaches for the Identification of Nuclear Transport Receptor Substrates 17. NPC Mimics: Probing the Mechanism of Nucleocytoplasmic Transport 18. Analysis of RNA Transport in Xenopus Oocytes and Mammalian Cells 19. Strategies for Investigating Nuclear-Cytoplasmic tRNA Dynamics in Yeast and Mammalian Cells 20. Dissecting Ribosome Assembly and Transport in Budding Yeast 21. Approaches to Studying Subnuclear Organization and Gene-Nuclear Pore Interactions
