Nuclear Structure and Function (Methods in Cell Biology) (Methods in Cell Biology)

This volume is a comprehensive guide to the methodologies used in the study of structural domains of cell nuclei. The text covers chromatin, the karyoskeleton, the soluble domain, and the nucleolus. It details methods that are used to isolate components from these domains and techniques used to assemble and disassemble nuclear elements. There is also coverage of three-dimensional mapping and localization of nuclear processes. Key Features\* Provides a practical laboratory guide for studying cell nuclei\* Includes comprehensive and easy-to-follow protocols Content: Series Editors Page ii Edited by Page iii Copyright Page Page iv Contributors Pages xiii-xv Preface Page xvii Miguel Berrios Chapter 1 Isolation and Characterization of Karyoskeletal Pro tein-Enriched Fractions from Vertebrate Livers Original Research Article Pages 3-22 Miguel Berrios Chapter 2 Preparation of Karyoskeletal Protein-Enriched Fractions from Drosophila melanogaster Cells and Tissues Original Research Article Pages 23-32 Paul A. Fisher Chapter 3 Isolation of Nuclei and Nucleoli from the Yeast Saccharomyces cerevisiae Original Research Article Pages 33-46 J.E. Dove, J.S. Brockenbrough, J.P. Aris Chapter 4 Determining Nuclear Structure Using the Fluorescence Light Microscope Original Research Article Pages 49-77 Michael R. Paddy Chapter 5 Localization of Single Nuclear Pore Complexes by Confocal Laser Scanning Microscopy and Analysis of Their Distribution Original Research Article Pages 79-98 Ulrich Kubitscheck, Reiner Peters Chapter 6 Nuclear Ultrastructure: Transmission Electron Microscopy and Image Analysis Original Research Article Pages 99-124 Andrew S. Belmont Chapter 7 Three-Dimensional Surface Structure Analysis of the Nucleus Original Research Article Pages 125-138 T.D. Allen, S.A. Rutherford, G.R. Bennion, C. Wiese, S. Riepert, E. Kiseleva, M.W. Goldberg Chapter 8 Scanning Transmission Electron Microscopy of Nuclear Structures Original Research Article Pages 139-164 Joseph S. Wall, James F. Hainfeld, Martha N. Simon Chapter 9 Electron Microscopic Imaging of Chromatin with Nucleosome Resolution Original Research Article Pages 167-186 C.L. Woodcock, R.A. Horowitz Chapter 10 Mapping Three-Dimensional Chromosome Architecture in Situ Original Research Article Pages 187-233 Abby F. Dernburg, John W. Sedat Chapter 11 Structural Analysis of Meiotic Chromosomes and Synaptonemal Complexes in Higher Vertebrates Original Research Article Pages 235-256 Alberto J. Solari Chapter 12 Genetic and Morphological Approaches for the Analysis of Meiotic Chromosomes in Yeast Original Research Article Pages 257-285 Josef Loidl, Franz Klein, Joanne Engebrecht Chapter 13 Mapping Proteins to Nuclear Pore Complexes by Immunogold Electron Microscopy Original Research Article Pages 287-302 Monika Grote, Reiner Peters Chapter 14 Methods Used to Study Structure and Function of the Nucleolus Original Research Article Pages 303-321 Robert L. Ochs Chapter 15 Identification of Base-Unpairing Region-Binding Proteins and Characterization of Their in Vivo Binding Sequences Original Research Article Pages 323-354 T. Kohwi-Shigematsu, I. Debelle, L.A. Dickinson, S. Galande, Y. Kohwi Chapter 16 Cell-Free Nuclear Reassembly in Mammalian Mitotic Homogenates Original Research Article Pages 357-366 Brian Burke Chapter 17 Analysis of Nuclear Envelope Assembly Using Extracts of Xenopus Eggs Original Research Article Pages 367-395 M.J. Lohka Chapter 18 Cell-Free Nuclear Assembly and Disassembly in Drosophila Original Research Article Pages 397-416 Paul A. Fisher, Miguel Berrios Chapter 19 Methods for Studying in Vitro Assembly of Male Pronuclei Using Oocyte Extracts from Marine Invertebrates Sea Urchins and Surf Clams Original Research Article Pages 417-452 Philippe Collas, Dominic Poccia Chapter 20 Mapping of DNA Replication Sites in Situ by Fluorescence Microscopy Original Research Article Pages 455-469 Roel van Driel, Erik M.M. Manders, Luitzen de Jong, Jan Stap, Jacob A. Aten Chapter 21 EM Visualization of Transcriptionally Active Genes after Injection into Xenopus Oocyte Nuclei Original Research Article Pages 471-496 Yvonne N. Osheim, Ann L. Beyer Chapter 22 Cell-Free Systems to Study Chromatin Remodeling Original Research Article Pages 497-515 Gregory H. Leno Chapter 23 In Vitro Systems for the Reconstitution of snRNP and Protein Nuclear Import Original Research Article Pages 517-543 Colin Dingwall, Isabel Palacios Chapter 24 In Vivo Nuclear Transport Kinetics in Saccharomyces cerevisiae Original Research Article Pages 545-557 Paul M. Roberts, David S. Goldfarb Chapter 25 Nuclear Transport of RNAs in Microinjected Xenopus Oocytes Original Research Article Pages 559-589 Michael P. Terns, David S. Goldfarb Chapter 26 In Vivo Systems to Study the Dynamics of Nuclear Lamins Original Research Article Pages 591-602 Ricardo Benavente, Georg Krohne Index Pages 603-619 Volumes In Series Pages 621-625 Part I. Cell Fractionation Preparation And Characterization Of Probes. 1. Isolation And Characterization Of Karyoskeletal Protein-enriched Fractions From Vertebrate Livers -- 2. Preparation Of Karyoskeletal Protein-enriched Fractions From Drosophila Melanogaster Cells And Tissues -- 3. Isolation Of Nuclei And Nucleoli From The Yeast Saccharomyces Cerevisiae -- Part Ii. Structural Analysis Of The Interphase Nucleus. 4. Determining Nuclear Structure Using The Fluorescence Light Microscope -- 5. Localization Of Single Nuclear Pore Complexes By Confocal Laser Scanning Microscopy And Analysis Of Their Distribution -- 6. Nuclear Ultrastructure: Transmission Electron Microscopy And Image Analysis -- 7. Three-dimensional Surface Structure Analysis Of The Nucleus -- 8. Scanning Transmission Electron Microscopy Of Nuclear Structures -- Part Iii. Chromatin And Subnuclear Structures. 9. Electron Microscopic Imaging Of Chromatin With Nucleosome Resolution -- 10. Mapping Three-dimensional Chromosome Architecture In Situ -- 11. Structural Analysis Of Meiotic Chromosomes And Synaptonemal Complexes In Higher Vertebrates -- 12. Genetic And Morphological Approaches For The Analysis Of Meiotic Chromosomes In Yeast -- 13. Mapping Proteins To Nuclear Pore Complexes By Immmunogold Electron Microscopy -- 14. Methods Used To Study Structure And Function Of The Nucleolus -- 15. Identification Of Base-unpairing Region-binding Proteins And Characterization Of Their In Vivo Binding Sequences -- Part Iv. Nuclear Assembly/disassembly In Cell-free Systems. 16. Cell-free Nuclear Reassembly In Mammalian Mitotic Homogenates -- 17. Analysis Of Nuclear Envelope Assembly Using Extracts Of Xenopus Eggs -- 18. Cell-free Nuclear Assembly And Disassembly In Drosophila -- 19. Methods For Studying In Vitro Assembly Of Male Pronuclei Using Oocyte Extracts From Marine Invertebrates: Sea Urchins And Surf Clams -- Part V. Localization Of Nuclear Processes. 20. Mapping Of Dna Replication Sites In Situ By Fluorescence Microscopy -- 21. Em Visualization Of Transcriptionally Active Genes After Injection Into Xenopus Oocyte Nuclei -- 22. Cell-free Systems To Study Chromatin Remodeling -- 23. In Vitro Systems For The Reconstitution Of Snrnp And Protein Nuclear Import -- 24. In Vivo Nuclear Transport Kinetics In Saccharomyces Cerevisiae -- 25. Nuclear Transport Of Rnas In Microinjected Xenopus Oocytes -- 26. In Vivo Systems To Study The Dynamics Of Nuclear Lamins. Edited By Miguel Berrios. Includes Bibliographical References And Index. This volume is a comprehensive guide to the methodologies used in the study of structural domains of cell nuclei. The text covers chromatin, the karyoskeleton, the soluble domain, and the nucleolus. It details methods that are used to isolate components from these domains and techniques used to assemble and disassemble nuclear elements. There is also coverage of three-dimensional mapping and localization of nuclear processes.

Key Features
* Provides a practical laboratory guide for studying cell nuclei
* Includes comprehensive and easy-to-follow protocols Many methods have been developed for the isolation and subfractionation of vertebrate liver nuclei to produce relatively insoluble karyoskeletal protein-enriched fractions [for reviews of the early literature, please see Franke (1974), Harris and Agutter (1976), Fry (1977), Harris (1977, 1978), and Agutter and Richardson (1980)].