Nuclear pore complex and nucleocytoplasmic transport - Methods: Methods in Cell Biology

This volume describes modern tools and techniques to study nuclear pore complexes and nucleocytoplasmic transport in diverse eukaryotic models systems (mammalian cells, Xenopus, C. elegans, yeast) The volume enables investigators to analyse 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 people studying 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 complex 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. This volume describes modern tools and techniques to study nuclear pore complexes and nucleocytoplasmic transport in diverse eukaryotic models systems (mammalian cells, Xenopus, C. elegans, yeast) Chapters are written by experts in the field Cutting-edge material INDICE: Imaging yeast NPCs: from classical Electron microscopy to Immuno SEM Imaging metazoan NPCs by FESEM Exploring nuclear pore complex structure and nucleocytoplasmic transport by immuno-electron microscopy using Xenopus oocyte nuclei Labeling of subunits and single proteins for electron microscopy by a molecular pointer: a tool to map NPC architecture. Determination of human nuclear pore complex stoichiometry by targeted proteomics Nuclear pore complex maintenance in long-lived cells. A pulse-chase epitope labeling to study cellular dynamics of newly synthesized proteins : a novel strategy to characterize NPC biogenesis and ribosome maturation/export Analyzing nuclear pore assembly in an in vitro nuclear reconstitution system Xenopus in vitro assays to analyze the function of TM Nups and targeting of INM proteins Imaging the assembly, structure and function of the nuclear pore inside cells Cell-fusion based assay to visualize interphase NPC formation in mammalian cells An in vitro system to study mitotic disassembly of nuclear pore complexes Modern tools to study nuclear pore complexes and nucleocytoplasmic transport in Caenorhabditis elegans Analysis of nucleocytoplasmic transport in digitonin-permeabilized mammalian cells Novel approaches for the identification of nuclear transport receptor substrates NPC mimics: probing the mechanism of nucleocytoplasmic transport Analysis of nuclear RNA export in Xenopus oocytes and mammalian cells Light sheet fluorescence microscopy for the study of nuclear export of native mRNPs Super-registration for studies of nucleocytoplasmic transport of mRNA In vivo strategies for investigating nuclear - cytoplasmic tRNA dynamics. Dissecting ribosome assembly and transport in budding yeast Microscopic and molecular approaches to studying gene-nuclear pore interactions Analysis of kinetochore-nuclear pore complex interactions during mitosis in yeast

• ISBN: 978-0-12-417160-2
• Editorial: Academic Press
• Encuadernacion: Cartoné
• Páginas: 464
• Fecha Publicación: 13/07/2014
• Nº Volúmenes: 1
• Idioma: Inglés