Description
Book SynopsisThis volume covers all methods used to discover the composition, structure, and dynamics of the nuclear pore complex (NPC), as well as the soluble transport factors involved in the transport process.
Table of ContentsPart I: The Nuclear Pore Complex and Nucleoporins
1. Affinity Isolation of Endogenous Saccharomyces cerevisiae Nuclear Pore Complexes
Ilona Nudelman, Javier Fernandez-Martinez, and Michael P. Rout
2. Transformation of Chaetomium thermophilum and affinity-purification of native thermostable protein complexes
Nikola Kellner and Ed Hurt
3. Nuclear Pore Complex Assembly using Xenopus Egg Extract
Guillaume Holzer and Wolfram Antonin
Part II: Nucleo-cytoplasmic Passage
4. Analysis of Nuclear Pore Complex Permeability in Mammalian Cells and Isolated Nuclei Using Fluorescent Dextrans
Marcela Raices and Maximiliano A. D’Angelo
5. Hormone-inducible Transport Reporter Assay to Study Nuclear Import Defects in Neurodegenerative Diseases
Saskia Hutten and Dorothee Dormann
6. Subcellular Fractionation Suitable for Studies of RNA and Protein Trafficking
Biljana Culjkovic-Kraljacic and Katherine L.B. Borden
7. Localising Total mRNA in Plant Cells
Geraint Parry
8. Using Single Molecule RNA FISH to Determine Nuclear Export and Transcription Phenotypes in Drosophila Tissues
Jennifer R. Aleman, Shawn C. Little, and Maya Capelson
Part III: Functional Analysis of Nucleoporins
9. Analysis of Nucleoporin Function using Inducible Degron Techniques
Vasilisa Aksenova, Alexei Arnaoutov, and Mary Dasso
10. Monitoring of Chromatin Organization at the Nuclear Pore Complex, Inner Nuclear Membrane and Nuclear Interior in Live Cells by Fluorescence Ratiometric Imaging of Chromatin (FRIC)
Frida Niss, Cecilia Bergqvist, Anna-Lena Ström, and Einar Hallberg
11. Analysis of Nuclear Pore Complexes in Caenorhabditis elegans by Live Imaging and Functional Genomic
Patricia de la Cruz Ruiz, Raquel Romero-Bueno, and Peter Askjaer
12. Protein Retargeting in Aspergillus nidulans to Study the Function of Nuclear Pore Complex Proteins
Subbulakshmi Suresh and Stephen A. Osmani
Part IV: Protein-Protein Interactions
13. Split-GFP Complementation to Study the Nuclear Membrane Proteome using Microscopy. Shary N. Shelton, Sarah E. Smith, and Sue L. Jaspersen
14. Bimolecular Fluorescence Complementation: Quantitative Analysis of In Cell Interaction of Nuclear Transporter Importin a with Cargo Proteins
Alexander Lee, Marie A. Bogoyevitch and David A. Jans
15. Validation of Nuclear Pore Complex Protein-Protein Interactions by Transient Expression in Plants
Fumika Ikeda and Kentaro Tamura
16. Binding affinity measurement of Nuclear Export Signal peptides to their exporter CRM1. Ho Yee Joyce Fung, and Yuh Min Chook
Part V: Post Translational Modifications
17. Analysis of Ubiquitylation and SUMOylation of Yeast Nuclear Pore Complex Proteins
Catherine Dargemont
18. Purification of Cdk-CyclinB-Kinase-Targeted Phosphopeptides from Nuclear Envelope
Justin D. Blethrow, Amanda L. DiGuilio, and Joseph S. Glavy
Part VI: Biophysical Methods
19. Crystallization of Nuclear Export Signals or Small Molecule Inhibitors Bound to Nuclear Exporter CRM1
Yee Joyce Fung and Yuh Min Chook
20. Atomic Force Microscopy for Structural and Biophysical Investigations on Nuclear Pore Complexes
Ivan Liashkovich, Gonzalo Rosso, and Victor Shahin
21. Multivalent Interactions with Intrinsically Disordered Proteins Measured by Surface Plasmon Resonance
Larisa E. Kapinos and Roderick Y. H. Lim
22. Assembly and Use of a Microfluidic Device to Study Nuclear Mechanobiology during Confined Migration
Richa Agrawal, Aaron Windsor, and Jan Lammerding
Part VII: Imaging NPCs and Transport
23. SPEED Microscopy: High-Speed Single-Molecule Tracking and Mapping of Nucleocytoplasmic Transport
Steven J. Schnell, Mark Tingey, and Weidong Yang
24. Imaging Fluorescent Nuclear Pore Complex Proteins in C. elegans
Courtney Lancaster, Giulia Zavagno, James Groombridge, Adelaide Raimundo, David Weinkove, Tim Hawkins, Joanne Robson and Martin W. Goldberg
25. Visualizing Nuclear Pore Complexes in Xenopus Egg Extracts
Sampada Mishra and Daniel L. Levy
26. TEM Imaging of Membrane Choreography During Mitosis of Drosophila Tissue Culture Cells
Anton Strunov, Lidiya V. Boldyreva, Alexey V. Pindyurin, Maurizio Gatti, and Elena Kiseleva
27. Scanning Electron Microscopy (SEM) and Immuno-SEM of Nuclear Pore Complexes from Amphibian Oocytes, Mammalian Cell Cultures, Yeast and Plants
Martin W. Goldberg and Jindřiška Fišerová
28. NPC Structure in Model Organisms: Transmission Electron Microscopy and Immuno-gold Labelling using High Pressure Freezing/Freeze Substitution of Yeast, Worms and Plants
A. Christine Richardson, Jindřiška Fišerová, and Martin W. Goldberg
29. High Resolution Imaging and Analysis of Individual Nuclear Pore Complexes
Boris Fichtman, Saroj G. Regmi, Mary Dasso, and Amnon Harel
30. Live CLEM imaging of Tetrahymena to Analyze the Dynamic Behavior of the Nuclear Pore Complex
Tokuko Haraguchi, Hiroko Osakada, and Masaaki Iwamoto
31. Visualizing Nuclear Pore Complex Assembly In Situ in Human Cells at Nanometer Resolution by Correlating Live Imaging with Electron Microscopy
Helena Bragulat-Teixidor, M. Julius Hossain, and Shotaro Otsuka
