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Author : Leena Gandhi
Publisher :
ISBN 13 :
Total Pages : 432 pages
Book Rating : 4.:/5 (34 download)
Download or read book Biochemical Characterization of Tetrahymena Thermophila Telomerase Proteins written by Leena Gandhi and published by . This book was released on 1998 with total page 432 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Author : Henry Chan
Publisher :
ISBN 13 :
Total Pages : 145 pages
Book Rating : 4.:/5 (14 download)
Download or read book Structural and Functional Characterization of the Tetrahymena Telomerase Holoenzyme written by Henry Chan and published by . This book was released on 2017 with total page 145 pages. Available in PDF, EPUB and Kindle. Book excerpt: This dissertation describes the structural characterization of the telomerase holoenzyme from the ciliate Tetrahymena thermophila using a combination of X-ray crystallography, cryogenic electron microscopy, and nuclear magnetic resonance spectroscopy. Telomerase is the ribonucleoprotein complex that iteratively reverse transcribes telomeric repeats to the ends of linear chromosomes in actively replicating eukaryotic cells. Previous biochemical and genetic studies of Tetrahymena telomerase holoenzyme identified the components TERT (the reverse transcriptase), TR (the internal RNA template), and six other accessory proteins (p65, Teb1, p50, p75, p45, and p19). Of these, p50, p75, p45, and p19 were largely uncharacterized and possessed no known homology to other proteins. A major focus of this work was to study p75-45-p19 which was known to form a trimeric complex. In the course of this work, crystal structures of p19 and the p45 C-terminal domain were solved and shown to be structurally homologous to the Ten1 and Stn1 proteins of the Replication Protein A (RPA)-like telomeric Ctc1-Stn1-Ten1 (CST) complex that is found in plants, vertebrates, and yeast, leading to the conclusion that p75-p45-p19 is Tetrahymena CST. In addition, the ongoing structural characterization of the p75 protein is discussed, in which a specific interaction between p75 and p50 is identified. The work in this thesis also contributed to the overall structural characterization of the entire holoenzyme through an electron microscopy (EM) collaboration with the Zhou lab where negative stain and cryogenic EM (cryo-EM) structures of Tetrahymena telomerase were solved to 25 and 9 i resolution respectively. These EM structures led to the characterization and unambiguous placement of all of the then known subunits including the placement of a homology model of the p75-p45-p19 trimeric core, based on the structure of p19 and the structure of the RPA trimeric core, into the cryo-EM structure, further confirming the characterization of p75-p45-p19 as Tetrahymena CST. The cryo-EM structure also led to the identification of two new subunits Teb2 and Teb3 by mass spectrometry, which were determined to form another trimeric RPA-like subcomplex, TEB, with Teb1. The identification and characterization of these TEB proteins led to the discovery that Teb2 and Teb3 are shared subunits with Tetrahymena RPA and of a new RPA-like protein, Rlp2, which forms an alternative complex with Teb3.
Author :
Publisher : Academic Press
ISBN 13 : 0080524958
Total Pages : 599 pages
Book Rating : 4.0/5 (85 download)
Download or read book Tetrahymena Thermophila written by and published by Academic Press. This book was released on 1999-09-08 with total page 599 pages. Available in PDF, EPUB and Kindle. Book excerpt: Tetrahymena thermophila is emerging as a powerful experimental system for functional genetics. The ciliated protozoan offers numerous advantages, not the least of which is the ability to eliminate any specific gene of interest and then to evaluate the effect of that mutation on the living cell. Past investigations with T. thermophila have yielded several key discoveries, including dynein, catalytic RNA, and telomerase. This volume is a comprehensive resource for using Tetrahymena in the laboratory and is aimed at persons already experienced, as well as newcomers to the organism. It covers both the biological background and essential protocols for investigators rapidly turning to Tetrahymena as the experimental system of choice. Contains both theoretical and practical issues in 30 chapters contributed by the world authorities on Tetrahymena Indispensible for both the novice and the experienced researcher Overviews the history, cell biology, and genetics of the organism Describes essential protocols on the growth of cells, genetic techniques, and how to look at the cells with the microscope Illustrates how the methods can be applied to solve various cell biological problems Reviews recently developed strategies for altering gene expression
Author : Stacey Cranert
Publisher :
ISBN 13 :
Total Pages : 136 pages
Book Rating : 4.:/5 (911 download)
Download or read book The Functional Evolution of Telomere Proteins in Tetrahymena Thermophila written by Stacey Cranert and published by . This book was released on 2014 with total page 136 pages. Available in PDF, EPUB and Kindle. Book excerpt: Telomeres are nucleo-protein complexes that contain DNA sequence complexed with specialized telomere binding proteins. The role of telomeres is to protect chromosome ends from being recognized as DNA damage and act as a buffer for DNA loss at the chromosome terminus. Telomeric DNA is a tandemly repeated sequence that consists of a double-stranded portion followed by a shorter, single-stranded overhang. Pot1 is a telomere binding protein that, in mammalian cells, binds the single stranded G-overhang of the telomere. It functions in telomere length regulation as well as in preventing the telomere from activating a cell cycle checkpoint. Tetrahymena thermophila have two POT homologs, Pot1 and Pot2. Pot1 is a functional homolog of the mammalian Pot1 in that it binds to the telomeric G-overhang and prevents it from activating a DNA damage response; however the function of Pot2 appears to be quite different. We have found that while Pot1 is essential during vegetative growth, Pot2 is not and Pot2 does not appear to have a role at macronuclear telomeres. RT-PCR and immunofluorescence analysis indicates that Pot2 is only expressed during the sexual phase of the Tetrahymena life cycle and it localizes to the developing macronucleus at a time when massive chromosome processing and new telomere formation occur. Chromatin immunoprecipitation (ChIP) assays demonstrate that Pot2 associates with sites of chromosome breakage (CBSs) but not sites of internal eliminated sequence (IES) excision during chromosome processing. In an effort to delineate the concurrent processes of chromosome breakage and IES excision, we analyzed the localization of the heterochromatic methylation marks, tri-methylated H3K9 and H3K27, during macronuclear development. It was known that these marks are essential for IES excision but their requirement for chromosome breakage was unclear. ChIP analysis shows that these marks are highly enriched at IESs but are not localized to CBSs. We also examined the requirement for expression of the essential IES excision protein Pdd1 in chromosome breakage. We confirm previous studies showing that parental expression of Pdd1 is not required for chromosome breakage, but demonstrate that complete loss of Pdd1 from both the parental and developing macronuclei results in ineffective chromosome breakage. Together, our results further support the evidence in the field that chromosome breakage and IES excision occur via different mechanisms but disruption of IES excision indirectly results in a defect in chromosome breakage. We have also begun the characterization of Pat2, a telomere protein that, unlike Pot2, functions at macronuclear telomeres in Tetrahymena. We show that knockout of Pat2 results in a telomerase-dependent decrease in telomere length without the rapid cell-cycle arrest seen with loss of Pot1. Co-immunoprecipitation analysis shows that Pat2 associates with the G-overhang binding complex in-vivo and this interaction is not dependent on DNA. These results indicate that Pat2 is a member of the G-overhang binding complex but it is unlikely to have a role in telomere protection. Instead Pat2 appears to positively regulate telomerase action at the telomere. The results presented in this dissertation demonstrate the evolution of two very different telomere protein homologs, Pot2 and Pat2. While Pat2 serves several canonical telomere protein functions, Pot2 has diverged to play other roles. Our analysis of Pot2 identifies the first protein in Tetrahymena to localize to CBSs and suggests that Pot2 could be involved in the exciting, and poorly characterized process of de novo telomere synthesis.
Author : Doreen Dianne Cunningham
Publisher :
ISBN 13 :
Total Pages : 310 pages
Book Rating : 4.:/5 (35 download)
Download or read book Biochemical and Biological Functions of Tetrahymena Thermophila Telomerase written by Doreen Dianne Cunningham and published by . This book was released on 2004 with total page 310 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Author : Bosun Min
Publisher :
ISBN 13 :
Total Pages : 226 pages
Book Rating : 4.:/5 (769 download)
Download or read book Telomerase Holoenzyme Proteins and Processivity Subunit in Tetrahymena Thermophila written by Bosun Min and published by . This book was released on 2009 with total page 226 pages. Available in PDF, EPUB and Kindle. Book excerpt: Telomeres are specialized protein-DNA structures that protect the ends of linear chromosomes, and they are maintained by the telomerase ribonucleoprotein (RNP) enzyme complex. Recombinant telomerase RNP with catalytic activity contains, at a minimum, the catalytic reverse transcriptase subunit (TERT) and the telomerase RNA (TER). However, endogenous telomerase is a much larger holoenzyme complex, with telomerase-associated subunits that contribute to RNP assembly and regulation. Telomerase-associated subunits may also directly affect the biochemical features of telomerase catalytic activity. In vitro reconstitution of Tetrahymena thermophila minimal RNP results in telomerase activity with low repeat addition processivity (RAP), while the endogenously assembled complexes can have both low and high RAP. Knowledge of a comprehensive list of telomerase-associated proteins expands our understanding of how telomerase-associated proteins regulate telomerase. Therefore, I optimized an affinity purification of TERT-associated proteins and identified telomerase-associated proteins to add to the list of known subunits. I have also begun characterizing the architecture of the telomerase holoenzyme: three subunits form a subcomplex that bridges the minimal RNP and a processivity factor, which contains telomeric single-stranded DNA-binding activity. With these telomerase subunits, a telomerase complex with high RAP can be reconstituted in vitro. The work described here expands the known functions of telomerase-associated proteins and the molecular mechanisms of telomere length regulation.
Author : Heather Upton
Publisher :
ISBN 13 :
Total Pages : 82 pages
Book Rating : 4.:/5 (957 download)
Download or read book Molecular Insights Into the Structure and Function of the Telomerase Holoenzyme in Tetrahymena Thermophila written by Heather Upton and published by . This book was released on 2016 with total page 82 pages. Available in PDF, EPUB and Kindle. Book excerpt: Telomeres are specialized, G-rich simple-sequence repeats that cap the ends of linear chromosomes to prevent genome instability. These tandem DNA repeats are bound by sequence-specific proteins to create a protective structure that marks the chromosome end thereby preventing aberrant chromosomal recombination, resection, degradation, and fusion. Due to inherent limitations of genome replication and chromosome end processing, telomeres shorten over time leading to potential loss of genetic information if not restored or maintained. The ribonucleoprotein (RNP) telomerase functions in this regard by using an integral RNA template (TER) to synthesize single stranded telomeric repeats at the chromosome end. In vitro minimal catalytic activity can be reconstituted from the telomerase protein component TERT and TER; however, in vivo biologically active holoenzyme requires further protein components for repeat addition synthesis, enzyme recruitment, and regulation in the cell. The ciliate Tetrahymena thermophila serves as an experimentally favorable model system for the study of telomerase due high levels of constitutively active enzyme and robust molecular and genetic techniques. Furthermore, our understanding of the holoenzyme is arguably best characterized from the Tetrahymena enzyme, which consists of nine protein components and the RNA (TERT, TER, p65, p50, Teb1, Teb2, Teb3, p75, p45, and p19). Despite knowledge of the overall architecture, relationships between multiple proteins within the holoenzyme and their specific physiological roles had remained unresolved. Using a variety of in vitro and in vivo biochemical techniques, I show that the holoenzyme component p50 functions as a central hub for enzyme assembly, connecting the RNP catalytic core to the RPA-like Teb1-Teb2-Teb3 (TEB) and p75-p45- p19 (CST) subcomplexes. To answer existing questions concerning telomerase recruitment, I employ endogenously tagged holoenzyme proteins to show that all telomerase holoenzyme subunits are subject to coordinate telomere recruitment and release dependent on the cell cycle. Using domain tagging and truncation strategies, I demonstrate that the high-affinity single-stranded telomeric DNA binding component Teb1 is necessary and sufficient for interaction between telomerase and the telomere. This work supports a model for Tetrahymena telomerase-telomere recruitment that breaks the precedent established by studies in yeast and vertebrate cells: Teb1- containing holoenzyme is recruited directly to the telomeric DNA rather than telomerase recruitment by interaction with a telomere-bound protein. Together, along with ongoing studies of the Tetrahymena TEB and CST subcomplexes, these results suggest commonalities of telomerase interaction, action, and regulation at telomeres across species.
Author : Hong Sheng
Publisher :
ISBN 13 :
Total Pages : 64 pages
Book Rating : 4.:/5 (324 download)
Download or read book Identification and Characterization of a Putative Telomere End-binding Protein from Tetrahymena Thermophila written by Hong Sheng and published by . This book was released on 1995 with total page 64 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Author : Ping Xie
Publisher :
ISBN 13 : 9781621009825
Total Pages : 0 pages
Book Rating : 4.0/5 (98 download)
Download or read book Tetrahymena and Human Telomerase Enzymes written by Ping Xie and published by . This book was released on 2012 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book gives a review of the author's previous work on modelling of the processive nucleotide and repeat addition translocations by recombinant Tetrahymena telomerase and proposes an analogous model for recombinant human telomerase. With the models, the dynamics of primer dissociation from the two enzymes and the dynamics of repeat addition translocation by the two enzymes are theoretically studied and compared in detail.
Author : Bruce Jon Mixan
Publisher :
ISBN 13 :
Total Pages : 90 pages
Book Rating : 4.:/5 (23 download)
Download or read book Isolation and Characterization of a G-strand Specific Telomere Binding Protein from Tetrahymena Thermophila written by Bruce Jon Mixan and published by . This book was released on 1989 with total page 90 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Author :
Publisher : Academic Press
ISBN 13 : 9780128163634
Total Pages : 0 pages
Book Rating : 4.1/5 (636 download)
Download or read book Transgenerational Epigenetics written by and published by Academic Press. This book was released on 2019-05-21 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Transgenerational Epigenetics, Second Edition, offers the only up-to-date, comprehensive analysis of the inheritance of epigenetic phenomena between generations with an emphasis on human disease relevance, drug discovery, and next steps in clinical translation. International experts discuss mechanisms of epigenetic inheritance, its expression in animal and plant models, and how human ailments, such as metabolic disorders and cardiovascular disease are influenced by transgenerational epigenetic inheritance. Where evidence is sufficient, epigenetic clinical interventions are proposed that may help prevent or reduce the severity of disease before offspring are born. This edition has been thoroughly revised in each disease area, featuring newly researched actors in epigenetic regulation, including long noncoding RNA in addition to histone modifications and DNA methylation. Therapeutic pathways in treating cancer and extending human longevity are also considered, as are current debates and future directions for research.
Author : Keiko Hiyama
Publisher : Springer Science & Business Media
ISBN 13 : 1603278796
Total Pages : 375 pages
Book Rating : 4.6/5 (32 download)
Download or read book Telomeres and Telomerase in Cancer written by Keiko Hiyama and published by Springer Science & Business Media. This book was released on 2009-03-18 with total page 375 pages. Available in PDF, EPUB and Kindle. Book excerpt: Telomerase, an enzyme that maintains telomeres and endows eukaryotic cells with immortality, was first discovered in tetrahymena in 1985. In 1990s, it was proven that this enzyme also plays a key role in the infinite proliferation of human cancer cells. Now telomere and telomerase are widely accepted as important factors involved in cancer biology, and as promising diagnostic tools and therapeutic targets. Recently, role of telomerase in “cancer stem cells” has become another attractive story. Until now, there are several good books on telomere and telomerase focusing on biology in ciliates, yeasts, and mouse or basic sciences in human, providing basic scientists or students with updated knowledge.
Author :
Publisher : Academic Press
ISBN 13 : 0128178205
Total Pages : 290 pages
Book Rating : 4.1/5 (281 download)
Download or read book Genome Plasticity in Health and Disease written by and published by Academic Press. This book was released on 2020-04-08 with total page 290 pages. Available in PDF, EPUB and Kindle. Book excerpt: Genome Plasticity in Health and Disease provides a fully up-to-date overview on genome plasticity and its role in human physiology and disease. Following an introduction to the field, a diverse range of chapters cover genomic and epigenomic analysis and the use of model organisms and genomic databases in studies. Specific molecular and biochemical mechanisms of genome plasticity are examined, including somatic variants, De Novo variants, founder variations, isolated populations dynamics, copy-number variations, mobile elements, DNA methylation, histone modifications, transcription factors, non-coding RNAs, telomere dynamics and RNA editing. Later chapters explore disease relevance for cancer, as well as cardiovascular, neuropsychiatric, inflammatory, and endocrine disease, and associated pathways for drug discovery. Examines the role of genome plasticity across a range of disease types, from cardiovascular disease, to cancer and neuropsychiatric disorders Adopts an interdisciplinary approach, with expert contributions across the spectrum of basic science and disease relevance to drug discovery
Author : Stephen Sonis
Publisher : Academic Press
ISBN 13 : 0128137630
Total Pages : 458 pages
Book Rating : 4.1/5 (281 download)
Download or read book Translational Systems Medicine and Oral Disease written by Stephen Sonis and published by Academic Press. This book was released on 2019-09-14 with total page 458 pages. Available in PDF, EPUB and Kindle. Book excerpt: Translational Systems Medicine and Oral Disease bridges the gap between discovery science and clinical oral medicine, providing opportunities for both the scientific and clinical communities to understand how to apply recent findings in cell biology, genomic profiling, and systems medicine to favorably impact the diagnosis, treatment and management of oral diseases. Fully illustrated chapters from leading international contributors explore clinical applications of genomics, proteomics, metabolomics, microbiomics and epigenetics, as well as analytic methods and functional omics in oral medicine. Disease specific chapters detail systems approaches to periodontal disease, salivary gland diseases, oral cancer, bone disease, and autoimmune disease, among others. In addition, the book emphasizes biological synergisms across disciplines and their translational impact for clinicians, researchers and students in the fields of dentistry, dermatology, gastroenterology, otolaryngology, oncology and primary care. Presents the work of leading international researchers and clinicians who speak on the clinical applications of genomics, proteomics, metabolomics, microbiomics, and epigenetics, as well as analytic methods and functional omics in oral medicine Provides full-color, richly illustrated chapters that examine systems approaches to periodontal disease, salivary gland diseases, oral cancer, bone disease and autoimmune diseases Includes clinical case studies that illustrate examples of oral disease diagnostics and management, highlighting points of key importance for the reader Emphasizes biological synergisms across disciplines and their translational impact for clinicians, researchers, and students in the fields of dentistry, dermatology, gastroenterology, otolaryngology, oncology, and primary care
Author : Denis Lynn
Publisher : Springer Science & Business Media
ISBN 13 : 1402082398
Total Pages : 627 pages
Book Rating : 4.4/5 (2 download)
Download or read book The Ciliated Protozoa written by Denis Lynn and published by Springer Science & Business Media. This book was released on 2008-06-24 with total page 627 pages. Available in PDF, EPUB and Kindle. Book excerpt: distances between groups of ciliates were as vast as significant hurdles to obtain copyright permissions the genetic distances between plants and animals for the over 1,000 required illustrations, and I put – THE major eukaryotic kingdoms at that time! the publication schedule ahead of this element. I continued to collaborate with Mitch, and in There are a number of significant illustrated guides 1991 my first “molecular” Magisterial student, to genera and species that have recently been pub- Spencer Greenwood, published an article estab- lished. References are made to these throughout lishing 1990 or thereabouts as the beginning of the book as sources that readers can consult for this the “Age of Refinement” – the period when gene aspect of ciliate diversity. A future project that I am sequencing techniques would deepen our under- contemplating is an illustrated guide to all the valid standing of the major lines of evolution within ciliate genera.
Author : Philip David Cohen
Publisher :
ISBN 13 :
Total Pages : 500 pages
Book Rating : 4.:/5 (33 download)
Download or read book Characterization of Telomeric Chromatin in Tetrahymena Thermophila written by Philip David Cohen and published by . This book was released on 1993 with total page 500 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Author : Hisao Masai
Publisher : Springer
ISBN 13 : 9811069557
Total Pages : 581 pages
Book Rating : 4.8/5 (11 download)
Download or read book DNA Replication written by Hisao Masai and published by Springer. This book was released on 2018-01-22 with total page 581 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book reviews the latest trends and future directions of DNA replication research. The contents reflect upon the principles that have been established through the genetic and enzymatic studies of bacterial, viral, and cellular replication during the past decades. The book begins with a historical overview of the studies on eukaryotic DNA replication by Professor Thomas Kelly, a pioneer of the field. The following chapters include genome-wide studies of replication origins and initiation factor binding, as well as the timing of DNA replications, mechanisms of initiation, DNA chain elongation and termination of DNA replication, the structural basis of functions of protein complexes responsible for execution of DNA replication, cell cycle-dependent regulation of DNA replication, the nature of replication stress and cells’ strategy to deal with the stress, and finally how all these phenomena are interconnected to genome instability and development of various diseases. By reviewing the existing concepts ranging from the old principles to the newest ideas, the book gives readers an opportunity to learn how the classical replication principles are now being modified and new concepts are being generated to explain how genome DNA replication is achieved with such high adaptability and plasticity. With the development of new methods including cryoelectron microscopy analyses of huge protein complexes, single molecular analyses of initiation and elongation of DNA replication, and total reconstitution of eukaryotic DNA replication with purified factors, the field is enjoying one of its most exciting moments, and this highly timely book conveys that excitement to all interested readers.
