Nuclear pore complexes and nucleocytoplasmic transport - Methods book cover

Nuclear pore complexes and nucleocytoplasmic transport - Methods

Methods in Cell Biology

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.

Audience

Researchers and students in cell, molecular and developmental biology

Hardbound, 554 Pages

Published: May 2014

Imprint: Academic Press

ISBN: 978-0-12-417160-2

Contents

  • 1. Fifty years of Nuclear Pores and Nucleocytoplasmic Transport Studies: Multiple Tools Revealing Complex Rules
    Aurélie Floch Benoit Palancade and Valérie Doye
    2. Imaging Metazoan Nuclear Pore Complexes by Field Emission Scanning Electron Microscopy
    Boris Fichtman, Lihi Shaulov, and Amnon Harel
    3. Imaging Yeast NPCs: From Classical Electron Microscopy to Immuno SEM
    Elena Kiseleva, A. Christine Richardson, Jindriska Fiserova, Anton A. Strunov, Matthew C. Spink, Simeon R. Johnson, and Martin W. Goldberg
    4. Exploring Nuclear Pore Complex Molecular Architecture by Immuno-Electron Microscopy using Xenopus Oocytes
    Nelly Panté and Birthe Fahrenkrog
    5. Utilizing the Dyn2 Dimerization-Zipper as a Tool to Probe NPC Structure and Function
    Dirk Flemming, Philipp Stelter, and Ed Hurt
    6. The use of Targeted Proteomics to Determine the Stoichiometry of Large Macromolecular Assemblies
    Alessandro Ori, Amparo Andres-Pons, and Martin Beck
    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
    Philipp Stelter and Ed Hurt
    8. Analysis of Nuclear Reconstitution, Nuclear Envelope Assembly, and Nuclear Pore Assembly using Xenopus in vitro Assays
    Cyril Bernis and Douglass J. Forbes
    9. Xenopus in vitro Assays to Analyze the Function of Transmembrane Nucleoporins and Targeting of Inner Nuclear Membrane Proteins
    Nathalie Eisenhardt, Allana Schooley, and Wolfram Antonin
    10. Imaging the Assembly, Structure and Function of the Nuclear Pore Inside Cells
    Shotaro Otsuka, Anna Szymborska, and Jan Ellenberg
    11. Cell-Fusion Method to Visualize Interphase Nuclear Pore (NPC) Formation
    Kazuhiro Maeshima, Tomoko Funakoshi, and Naoko Imamoto
    12. An in vitro System to Study Nuclear Envelope Breakdown
    Joseph Marino, Lysie Champion, Cornelia Wandke, Peter Horvath, Monika I. Mayr, and Ulrike Kutay
    13. Modern Tools to Study Nuclear Pore Complexes and Nucleocytoplasmic Transport in Caenorhabditis elegans
    Peter Askjaer, Vincent Galy, and Peter Meister
    14. Assessing Regulated Nuclear Transport in Saccharomyces cerevisiae
    Christopher Ptak and Richard W. Wozniak
    15. Analysis of Nucleocytoplasmic Transport in Digitonin-Permeabilized Cells under Different Cellular Conditions
    Maiko Furuta, Shingo Kose, Ralph H. Kehlenbach, and Naoko Imamoto
    16. Novel Approaches for the Identification of Nuclear Transport Receptor Substrates
    Makoto Kimura, Ketan Thakar, Samir Karaca, Naoko Imamoto, and Ralph H. Kehlenbach
    17. NPC Mimics: Probing the Mechanism of Nucleocytoplasmic Transport
    Tijana Jovanovic-Talisman, Brian T. Chait, and Michael P. Rout
    18. Analysis of RNA Transport in Xenopus Oocytes and Mammalian Cells
    Ichiro Taniguchi, Asako McCloskey, and Mutsuhito Ohno
    19. Strategies for Investigating Nuclear-Cytoplasmic tRNA Dynamics in Yeast and Mammalian Cells
    Jacqueline B. Pierce, Shawn C. Chafe, Manoja, B. K. Eswara, George van der Merwe, and Dev Mangroo
    20. Dissecting Ribosome Assembly and Transport in Budding Yeast
    Martin Altvater, Sabina Schütz, Yiming Chang, and Vikram Govind Panse
    21. Approaches to Studying Subnuclear Organization and Gene-Nuclear Pore Interactions
    Defne Emel Egecioglu, Agustina D’Urso, Donna Garvey Brickner, William H. Light, and Jason H. Brickner

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