Cell Cycle Control And Dysregulation Protocols

Cell Cycle Control and Dysregulation Protocols focuses on emerging methodologies for studying the cell cycle, kinases, and kinase inhibitors. It addresses the issue of gene expression in vivo and in vitro, the analysis of cyclin-dependent kinase inhibitors, protein degradation mediated by the proteosome, the analysis of the transformed cell phenotype, and innovative techniques to detect apoptosis. Because there are already many manuals and protocols available, along with commercial kits and reagents, a variety of the more common techniques have not been included in our book. The protocols described, based on rather sophisticated techniques for in vivo and in vitro studies, consist of molecular biology, biochemistry, and various types of immunoassays. Indeed, the authors have successfully accomplished an arduous task by presenting several topics in the simplest possible manner. We are confident that Cell Cycle Control and Dysregulation Protocols will facilitate and optimize the work of practical scientists involved in researching the cell cycle. We greatly acknowledge the extraordinary contribution of the authors in writing this book.

The field of cell cycle regulation is based on the observation that the life cycle of a cell progresses through several distinct phases, G1, M, S, and G2, occurring in a well-defined temporal order. Details of the mechanisms involved are rapidly emerging and appear extraordinarily complex. Furthermore, not only is the order of the phases important, but in normal eukaryotic cells one phase will not begin unless the prior phase is completed successfully. Che- point control mechanisms are essentially surveillance systems that monitor the events in each phase, and assure that the cell does not progress prematurely to the next phase. If conditions are such that the cell is not ready to progress—for example, because of incomplete DNA replication in S or DNA damage that may interfere with chromosome segregation in M—a transient delay in cell cycle progression will occur. Once the inducing event is properly handled— for example, DNA replication is no longer blocked or damaged DNA is repaired—cell cycle progression continues. Checkpoint controls have recently been the focus of intense study by investigators interested in mechanisms that regulate the cell cycle. Furthermore, the relationship between checkpoint c- trol and carcinogenesis has additionally enhanced interest in these cell cycle regulatory pathways. It is clear that cancer cells often lack these checkpoints and exhibit genomic instability as a result. Moreover, several tumor suppressor genes participate in checkpoint control, and alterations in these genes are as- ciated with genomic instability as well as the development of cancer.

A collection of new reviews and protocols from leading experts in cell cycle regulation, Cell Cycle Control: Mechanisms and Protocols, Second Edition presents a comprehensive guide to recent technical and theoretical advancements in the field. Beginning with the overviews of various cell cycle regulations, this title presents the most current protocols and state-of-the-art techniques used to generate latest findings in cell cycle regulation, such as protocols to analyze cell cycle events and molecules. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Cell Cycle Control: Mechanisms and Protocols, Second Edition will be a valuable resource for a wide audience, ranging from the experienced cell cycle researchers looking for new approaches to the junior graduate students giving their first steps in cell cycle research.

Cell cycle checkpoints control the fidelity and orderly progression of eukaryotic cell division. By controlling the orderly progression of critical cell cycle events such as DNA replication and chromosome segregation and ensuring proper repair of damaged DNA, cell cycle checkpoints function to ensure genome integrity. Mechanisms of checkpoint controls are not only the research focus of investigators interested in mechanisms that regulate the cell cycle, but are also the interests of researchers studying cancer development as it is increasingly clear that loss of cell cycle checkpoints, which leads to genomic instability as a result, is a hallmark of tumorigenesis. Cell Cycle Checkpoints: Methods and Protocols provides detailed descriptions of methodologies currently employed by researchers in the field, including those commonly used in the mammalian, yeast, C. elegans, Drosophila, and Xenopus model systems. Each chapter describes a specific technique or protocol, such as a method to induce cell cycle checkpoints in a particular model system, to synchronize a population of cells to allow observations of cell cycle progression, to identify genes involved in checkpoint regulation, and to study particular protein components of cell cycle checkpoint pathways. Written in the highly successful Methods in Molecular BiologyTM series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Cell Cycle Checkpoints: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies in an effort to further our knowledge of this essential field.

This volume covers a broad range of cell types including cultured cell lines, primary cells, and various unicellular organisms such as fission yeast, budding yeast, parasite Leishmania amazonensis, and parasite Trypanosoma brucei. The chapters in this book are organized into four parts. Part One looks at a general overview of cell cycle control and synchronization. Part Two discusses techniques to synchronize mammalian cells to various cell cycle phases including mitotic sub-phases. Part Three covers synchronization of unicellular organisms and Part Four analyzes cell cycle progression. Written in the highly successful Methods in Molecular Biology 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. Cutting-edge and thorough, Cell-Cycle Synchronization: Methods and Protocols is a valuable resource for both novice and expert scientists in this developing field.

This book brings together a unique collection of protocols that cover novel and specialized techniques as well as updated and improved adaptations of more standard procedures involving the cell cycle and its regulation by oscillatory networks, exploring recent progress in the field from both holistic and reductionist perspectives. The edition provides a space for researchers to highlight and explore the latest developments in molecular biology and biochemical techniques for studying oscillatory networks and to share these across the research community to facilitate further progress. Written in the highly successful Methods in Molecular Biology 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.Authoritative and up-to-date, Cell Cycle Oscillators: Methods and Protocols, Second Edition serves as an invaluable guide for those new to the field as well as the more experienced scientist. Chapters 19 and 21 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.