[PDF] Mutational Analysis Of Gamma Tubulin eBook

Author : Jason Harris
Publisher :
Page : pages
File Size : 17,19 MB
Release : 2015
Category :
ISBN :

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"Inaccurate segregation of chromosomes during cell division is thought to lead to tumour formation. Microtubules play a key role in chromosome separation and segregation. The study of yeast spindles offers a convenient and versatile system to study the processes that impact cell division. One protein found to have an integral role in proper spindle assembly is yeast [gamma]-Tubulin (Tub4). Tub4 is a 53kDa protein that contains a flexible C-terminal tail. Removal of this tail has shown to be lethal in vivo. Mutation of a DYSL motif in the tail has also been shown to influence recruitment of proteins to the opposite end of a microtubule. Phosphomimetic mutations of the tyrosine in the DYSL motif have been shown to increase the number of microtubules that do not attach to kinetochores. NMR is ideally suited to study the effects of phosphorylation on the dynamics of the last 39 residues of Tub4. In doing so, we will gain information on how changes in the dynamics of Tub4 correlate to changes in protein function, which in turn influence the cell as a whole.We have used NMR spectroscopy to characterize the differences between the wild type(WT) Tub4 tail and a mutant containing the Y445D phosphomimetic substitution. Gradient diffusion experiments were employed to measure the hydrodynamic radii of the proteins, revealing that both WT and Y445D constructs diffuse more slowly than expected. The dynamics of the two constructs were analyzed using 15N and 2H spin relaxation measurements. Spectral density mapping and analysis of backbone dynamics show no change in the fast timescale dynamics by the Y445D mutation. Interestingly, CPMG analysis revealed that the Y445D mutation changed the milli-microsecond timescale dynamics of the protein. This change is observed throughout the entire protein and was determined to be a global exchange between a major and minor state. This suggests that the mutation causes the protein to exchange between collapsed and extended conformations. In this model, the extended conformation increases the capture radius in which it can interact with potential binding partners." --

Author : Anne Straube
Publisher : Humana Press
Page : 319 pages
File Size : 26,60 MB
Release : 2017-04-30
Category : Science
ISBN : 9781493961856

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Microtubules are at the heart of cellular self-organization, and their dynamic nature allows them to explore the intracellular space and mediate the transport of cargoes from the nucleus to the outer edges of the cell and back. In Microtubule Dynamics: Methods and Protocols, experts in the field provide an up-to-date collection of methods and approaches that are used to investigate microtubule dynamics in vitro and in cells. Beginning with the question of how to analyze microtubule dynamics, the volume continues with detailed descriptions of how to isolate tubulin from different sources and with different posttranslational modifications, methods used to study microtubule dynamics and microtubule interactions in vitro, techniques to investigate the ultrastructure of microtubules and associated proteins, assays to study microtubule nucleation, turnover, and force production in cells, as well as approaches to isolate novel microtubule-associated proteins and their interacting proteins. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Definitive and practical, Microtubule Dynamics: Methods and Protocols provides the key protocols needed by novices and experts on how to perform a broad range of well-established and newly-emerging techniques in this vital field.

Author : John J. Correia
Publisher : Academic Press
Page : 472 pages
File Size : 13,69 MB
Release : 2013-08-20
Category : Science
ISBN : 0124078885

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There continues to be intense interest in the microtubule cytoskeleton; the assembly, structure and regulation of microtubules; and the numerous motors and accessory proteins that control cell cycle, dynamics, organization and transport. The field continues to grow and explore new aspects of these issues driven immensely by developments in optical imaging and tracking techniques. This Second Edition brings together current research and protocols in the field of microtubules in vitro and will serve as a valuable tool for cell biologists, biophysicists and pharmacologists who study the microtubule cytoskeleton, as well as for researchers in the biomedical and biotechnology communities with interest in developing drugs that target microtubules, MAPS and motors. Chapters reflect experimental procedures and new developments in the field of microtubule in vitro research Combines classical approaches and modern technologies Presents easy-to-use protocols and thorough background information, compiled by leaders in the field

Author :
Publisher : Academic Press
Page : 549 pages
File Size : 42,5 MB
Release : 2010-09-24
Category : Science
ISBN : 0123813506

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Microtubules: in vivo includes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton. The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation. Includes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation

Author :
Publisher : Elsevier
Page : 511 pages
File Size : 45,90 MB
Release : 2000-09-26
Category : Science
ISBN : 0080488625

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Centrosomes play an integral role in the growth of cells and the ultimate development of many animals, and sometimes plants. In addition to the normal growth of cells, centrosomes can also play a key role in the spread of cancer and are of increased interest to both the genetics and oncology communities. Volume 49 of Current Topics in Developmental Biology will present all known research surrounding the centrosome, across a variety of systems, will be well referenced, and speculate where the research is headed. Discusses centrosomes and cancer, centrosomes and early development, and molecular biology of the centrosome Heavily illustrated, with many color figures Chapters written by international leaders in the field

Author : Stavros J. Baloyannis
Publisher :
Page : 0 pages
File Size : 11,1 MB
Release : 2021
Category : Brain
ISBN : 9781839680984

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Cerebral and Cerebellar Cortex - Interaction and Dynamics in Health and Disease discusses several important issues of cerebro-cerebellar collaboration and interactions. The morphological and functional study of the cerebral and cerebellar cortices and their interaction has considerable value for interpreting the clinical phenomenology of cortical degenerations in the initial stage of the disease. In addition, the analysis of cerebro-cerebellar interactions strongly supports the concept of the close functional unity and harmonization of the brain and the cerebellum, underlining the important role that the cerebellar cortex plays in the performance of higher mental faculties, creativity, emotional processes, and homeostatic equilibrium of the human body.