Chemical Genetic Analysis of Signaling by the Saccharomyces Cerevisiae Mitotic Kinases Cdc15, Dbf2, and Cdc5 PDF Download
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Author: Jennifer L. Paulson
Publisher:
ISBN:
Category :
Languages : en
Pages : 292
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Book Description
Protein phosphorylation is a ubiquitous regulatory mechanism for cellular signal propagation, and the complexity of signaling networks presents a challenge to protein kinase substrate identification. Chemical genetic control of kinase function provides a handle for kinase pathway analysis. Here, we apply this approach to three kinases that function in a signaling network that regulates exit from mitosis in the budding yeast, Saccharomyces cerevisiae. These include the mitogen-activating protein kinase, Cdc15, the nuclear Dbf2-related kinase, Dbf2, and the Polo-like kinase, Cdc5. Each kinase was successfully engineered for selective chemical inhibition in vivo. We found that monospecific pharmacological inhibition of Cdc5 delays anaphase nucleus migration into the bud, revealing a novel Cdc5 function. Additionally, chemical genetic, bioinformatic, and yeast proteomic tools were combined for Cdc5 substrate identification. Systematically chosen candidate Cdc5 substrates were examined for loss of phosphorylation upon cellular Cdc5 inhibition. The identified Cdc5 targets include Spc72, a spindle pole body (SPB) component and microtubule anchor required for nuclear positioning. Spc72 binds Cdc5 in a cell cycle specific manner, and in vivo Cdc5 inhibition prevents mitotic Spc72 phosphorylation. Studies in vitro demonstrate direct Spc72 phosphorylation by Cdc5. Finally, we expanded our knowledge of Cdc5 function at the SPB by examining SPB-localized proteins for presence in a Cdc5 complex. In summary, a chemical genetic approach was used to inhibit three protein kinases from diverse families, which led to a greater understanding of Cdc5 cellular function.
Author: Jennifer L. Paulson
Publisher:
ISBN:
Category :
Languages : en
Pages : 292
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Book Description
Protein phosphorylation is a ubiquitous regulatory mechanism for cellular signal propagation, and the complexity of signaling networks presents a challenge to protein kinase substrate identification. Chemical genetic control of kinase function provides a handle for kinase pathway analysis. Here, we apply this approach to three kinases that function in a signaling network that regulates exit from mitosis in the budding yeast, Saccharomyces cerevisiae. These include the mitogen-activating protein kinase, Cdc15, the nuclear Dbf2-related kinase, Dbf2, and the Polo-like kinase, Cdc5. Each kinase was successfully engineered for selective chemical inhibition in vivo. We found that monospecific pharmacological inhibition of Cdc5 delays anaphase nucleus migration into the bud, revealing a novel Cdc5 function. Additionally, chemical genetic, bioinformatic, and yeast proteomic tools were combined for Cdc5 substrate identification. Systematically chosen candidate Cdc5 substrates were examined for loss of phosphorylation upon cellular Cdc5 inhibition. The identified Cdc5 targets include Spc72, a spindle pole body (SPB) component and microtubule anchor required for nuclear positioning. Spc72 binds Cdc5 in a cell cycle specific manner, and in vivo Cdc5 inhibition prevents mitotic Spc72 phosphorylation. Studies in vitro demonstrate direct Spc72 phosphorylation by Cdc5. Finally, we expanded our knowledge of Cdc5 function at the SPB by examining SPB-localized proteins for presence in a Cdc5 complex. In summary, a chemical genetic approach was used to inhibit three protein kinases from diverse families, which led to a greater understanding of Cdc5 cellular function.
Author:
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ISBN:
Category : Dissertations, Academic
Languages : en
Pages : 924
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Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 1508
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Book Description
Vols. for 1963- include as pt. 2 of the Jan. issue: Medical subject headings.
Author: Christine Guthrie
Publisher:
ISBN: 9780123106704
Category : Molecular biology
Languages : en
Pages : 933
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Book Description
Guide to Yeast Genetics and Molecular Biology presents, for the first time, a comprehensive compilation of the protocols and procedures that have made Saccharomyces cerevisiae such a facile system for all researchers in molecular and cell biology. Whether you are an established yeast biologist or a newcomer to the field, this volume contains all the up-to-date methods you will need to study "Your Favorite Gene" in yeast. Basic Methods in Yeast Genetics**Physical and genetic mapping**Making and recovering mutants**Cloning and Recombinant DNA Methods**High-efficiency transformation**Preparation of yeast artificial chromosome vectors**Basic Methods of Cell Biology**Immunomicroscopy**Protein targeting assays**Biochemistry of Gene Expression**Vectors for regulated expression**Isolation of labeled and unlabeled DNA, RNA, and protein
Author:
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ISBN:
Category : Cellular control mechanisms
Languages : en
Pages : 564
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Author: S. Guidet
Publisher: Springer Science & Business Media
ISBN: 1461518091
Category : Medical
Languages : en
Pages : 365
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Book Description
Progress in Cell Cycle Research is a new annual series designed to be the source for up-to-date research on this rapidly expanding field. Review articles by international experts examine various aspects of cell division regulation from fundamental perspectives to potential medical applications. Researchers as well as advanced undergraduate and graduate students in cell biology, biochemistry, and molecular biology will benefit from this series.
Author: David Owen Morgan
Publisher: New Science Press
ISBN: 0878935088
Category : Science
Languages : en
Pages : 328
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Book Description
The Cell Cycle: Principles of Control provides an engaging insight into the process of cell division, bringing to the student a much-needed synthesis of a subject entering a period of unprecedented growth as an understanding of the molecular mechanisms underlying cell division are revealed.
Author: Peter A. Hall
Publisher: John Wiley & Sons
ISBN: 9780470779699
Category : Science
Languages : en
Pages : 380
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Book Description
"The authors represent most of the key figures and the work and the book as a whole is an essential reference for the newcomer or specialist in this area and for any student of eukaryotic cell structure and function. This is an important and wonderful reference." –Microbiology Today, May 2009 Septins are an evolutionarily conserved group of GTP-binding and filament-forming proteins that were originally discovered in yeast. Once the preserve of a small band of yeast biologists, the field has grown rapidly in the past few years and now encompasses the whole of animal and fungal biology. Furthermore, septins are nowadays recognized to be involved in a variety of disease processes from neoplasia to neurodegenerative conditions. This book comprehensively examines the septin gene family and their proteins, providing those new to this research area with a detailed and wide ranging introduction to septin biology. It starts with a unique historical perspective on the development of the field, from its beginnings in the screen for cell division mutants by the Nobel Laureate Lee Hartwell. The evolution of the septin gene family then forms a basis for consideration of the biochemistry and functions of septins in yeast and other model organisms including C. elegans and Drosophila. A major part of the book considers the diversity of septins in mammals, their functions and properties as well as their involvement in normal and abnormal cellular states, followed by a speculative overview from the editors of the key questions in septin research and of where the field may be headed. In addition, several appendices summarise important information for those in, or just entering, the field, e.g. nomenclature and septin and septin-like sequences. This book is an essential source of reference material for researchers in septin biology, cell biology, genetics and medicine, in particular pathology, including areas of neurobiology, oncology, infectious disease and developmental biology.
Author:
Publisher:
ISBN:
Category : Biomolecules
Languages : en
Pages : 1006
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Author: Karl Esser
Publisher: Springer Science & Business Media
ISBN:
Category : Health & Fitness
Languages : en
Pages : 480
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Book Description
Mycology, the study of fungi, originated as a subdiscipline of botany and was a descriptive discipline, largely neglected as an experimental science until the early years of this century. A seminal paper by Blakeslee in 1904 provided evidence for self-incompatibility, termed "heterothallism", and stimulated interest in studies related to the control of sexual reproduction in fungi by mating-type specificities. Soon to follow was the demonstration that sexually reproducing fungi exhibit Mendelian inheritance and that it was possible to conduct formal genetic analysis with fungi. The names Burgeff, Kniep and Lindegren are all associated with this early period of fungal genetics research. These studies and the discovery of penicillin by Fleming, who shared a Nobel Prize in 1945, provided further impetus for experimental research with fungi. Thus began a period of interest in mutation induction and analysis of mutants for bio chemical traits. Such fundamental research, conducted largely with Neurospora crassa, led to the one gene: one enzyme hypothesis and to a second Nobel Prize for fungal research awarded to Beadle and Tatum in 1958. Fundamental research in biochemical genetics was extended to other fungi, especially to Saccharomyces cerevisiae, and by the mid-1960s fungal systems were much favored for studies in eukaryotic molecular biology and were soon able to compete with bacterial systems in the molecular arena.
