Cell and Molecular Biology of the Cytoskeleton

1 Probing the Cytoskeleton by Microinjection.- 1. Introduction.- 2. The Microtubule System.- 2.1. Analyzing the Equilibrium between Tubulin and Microtubules.- 2.2. Studying Microtubular Dynamics.- 2.3. Analyzing Microtubule-Bound Organelle Transport.- 2.4. Introducing Antitubulins.- 3. The Microfilament System.- 3.1. Studying Actin Dynamics.- 3.2. Studying the Dynamics of Actin-Associated Proteins.- 3.3. Reorganizing Microfilament Patterns by Introducing Structural Proteins.- 3.4. Introducing Phalloidin.- 3.5. Using Antibodies against Microfilament Proteins.- 4. The Intermediate Filament System.- 4.1. Introducing Antibodies.- 4.2. Introducing mRNA.- 5. The Coated Vesicle System.- 6. Conclusion.- References.- 2 Structural Aspects of Intermediate Filaments.- 1. Introduction.- 2. Domain Organization of IF Proteins.- 3. Neurofilaments.- 4. Organization of the Sequences along the Rod Domain.- 5. Comparison of the Six Nonepithelial Proteins along the Rod Domain.- 6. Structural Characterization of the Rod.- 7. From the Rodlike Protofilament to the 10-nm Filament.- 8. Structure and Function of the Terminal Domains in IF Proteins.- 8.1. The Involvement of Headpieces in Filament Formation.- 8.2. Structural Characterization of the Tailpieces.- 8.3. Are the Carboxyterminal Tailpieces Appendages Communicating between Stiff Filaments and the Cytoplasm?.- 9. Addendum.- References.- 3 The Structure and Evolution of Intermediate Filament Genes.- 1. Introduction.- 1.1. Correlation of Keratin Gene Expression with Differentiation State.- 1.2. Correlation of Keratin Subunit Structure with Differential Expression.- 2. Structure of Intermediate Filament Genes.- 3. A Model for the Evolution of Intermediate Filament Genes.- 3.1. Evolution of the Rod Domains of Intermediate Filament Genes.- 3.2. Evolution of the End Domains.- 4. Conclusion.- References.- 4 Differential Expression of the Genes Encoding the Keratins of Cultured Human Epidermal Cells.- 1.Introduction.- 2.The Structure of the 8-nm Keratin Filaments of Epidermis.- 3. The Genes Encoding the Human Epidermal Keratins.- 4. Structure of the Two Types of Keratin Genes and Their Relation to Other Intermediate-Filament Genes.- 5. Regulation of the Type I and Type II Keratin Genes.- References.- 5 Organization and Expression of the Vimentin and Desmin Genes.- 1. The Discovery of Intermediate Filaments.- 2. The Diversity of IF Subunits.- 3. The Genes Encoding IF.- 4. Expression of the Cloned Desmin Gene.- 4.1. In Lens Cells.- 4.2. Expression of the Hamster Desmin Gene in Human Epithelial (HeLa) Cells.- 5. Chromosome Localization of the Human Vimentin and Desmin Gene.- References.- 6 Actin: A Regulator of Cell Growth and Differentiation.- 1. Introduction.- 2. Cell Morphology and Growth and Differentiation.- 2.1. Cell Configuration.- 2.2. Cytoskeletal Organization.- 3. Regulation of Actin Gene Expression.- 4. Is Actin a Protooncogene?.- 5. Mechanisms by Which Actin May Regulate Cell Growth and Differentiation.- References.- 7 The Actin Genes in Caenorhabditis elegans.- 1. Introduction.- 2. Genomic Organization.- 3. Sequence Analysis.- 4. Transcription.- 5. Translational Products.- 6. Identification of Actin Mutants.- 7. Actin Gene Organization in Revertants of Actin Mutants.- 8. Transcription in Revertants.- 9. Future Prospects.- References.- 8 Implications of Microtubule Polarity for Microtubule Function.- 1. Introduction.- 2. Background and Methods for Determining MT Polarity.- 3. MT Polarity and Spindle Function.- 4. MT Polarity and Intracellular Motility.- References.- 9 Molecular Mechanisms Controlling Tubulin Synthesis.- 1. Introduction.- 2. Apparent Autoregulatory Control of Tubulin Synthesis in Animal Cells.- 2.1. Tubulin Synthesis in Animal Cells Is Sensitive to the Apparent Pool of Unpolymerized Subunits.- 2.2. Possible Molecular Levels for Regulating Tubulin Synthetic Rates.- 2.3. Transient Induction of Tubulin Synthesis by Interferon.- 3. Programmed Synthesis of Tubulin during Flagellar Growth in Unicellular Organisms.- 3.1. Regulated Synthesis of Tubulin Achieved through a Combination of Transcriptional Control and mRNA Stability in Chlamydomonas.- 3.2. Identification of Candidate DNA Sequences That Mediate Tubulin Regulation in Chlamydomonas.- 3.3. The Paradigm of Tubulin Regulation in Naegleria.- 3.4. Altered Tubulin Regulation in the Life Form Switch of the Parasitic Protozoa Leishmania.- 4. Programmed Synthesis of Tubulin during the Cell Cycle.- 4.1. The Synchronous Cell Cycle of Physarum.- 4.2. Cell Cycle Regulation in Other Eukaryotes.- 5. Summary.- References.- 10 Regulation of Tubulin Expression in Brain.- 1. Introduction.- 2. Tubulin Proteins.- 2.1. Different Isoforms of ?- and ?-Tubulins.- 2.2. Differential Localization of Brain ?- and ?-Tubulin Isoforms.- 2.3. Evidence for Posttranslational Modification.- 2.4. Developmental Regulation of Tubulin Isoforms.- 3. Tubulin Genes in Higher Eukaryotes.- 3.1. ?-Tubulin Genes.- 3.2. ?-Tubulin Genes.- 4. Expression of ?-Tubulin mRNAs.- 4.1. In Vitro Translation Products of ?-Tubulin mRNAs.- 4.2. Hybridization Analysis of Different ?-Tubulin mRNAs.- 5.Expression of ?-Tubulin mRNAs.- 5.1. In Vitro Translation Products of ?-Tubulin mRNAs.- 5.2. Hybridization Analysis of ?-Tubulin mRNAs.- 5.3. In Situ Hybridization of Chick ?2-Tubulin cDNA in Rat Cerebellum.- 6. Conclusions.- References.- 11 Tubulins from Plants, Fungi, and Protists: A Review.- 1. Introduction: Scope and Objectives.- 2. Isolation and Identification of Tubulin.- 2.1. Assays for Tubulin.- 2.2. Ion-Exchange Chromatography.- 2.3. Drug Affinity Methods.- 2.4. Microtubule Polymerization and Depolymerization.- 2.5. Copolymerization with Heterologous Tubulins.- 2.6. Tubulin-Rich Organelles.- 2.7. Technical Problems.- 3. Tubulin Dimer Structure.- 3.1. Amino Acid Sequences.- 3.2. Electrophoretic Resolution of ? and ? Subunits.- 3.3. Peptide Mapping Patterns.- 3.4. Immunology.- 3.5. Subunit Microheterogeneity.- 3.6. Posttranslational Modifications.- 3.7. Multiple Tubulin Genes.- 3.8. Membrane-Associated Tubulin.- 4. Antimicrotubule Drugs.- 4.1. Colchicine Effects in Vivo.- 4.2. Colchicine Effects in Vitro.- 4.3. Antimicrotubule Herbicides.- 4.4. Benzimidazole Derivatives.- 4.5. Taxol.- 4.6. Other Agents.- 5. Summary and Future Directions.- References.