Control of MRNA Metabolism by Poly(A) Binding Protein and Development of the Tethered Function Assay PDF Download
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Author: Jeffery Michael Coller
Publisher:
ISBN:
Category :
Languages : en
Pages : 182
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Author: Jeffery Michael Coller
Publisher:
ISBN:
Category :
Languages : en
Pages : 182
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Author: Zdravko Lorkovic
Publisher: CRC Press
ISBN: 149871336X
Category : Science
Languages : en
Pages : 174
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Book Description
Gene expression in eukaryotes is regulated at different levels, which need to be coordinated to implement the information in the genome. Now it is clear that post-transcriptional regulation of gene expression such as pre-mRNA splicing, mRNA transport, editing, turnover and translation are as important as the control of transcription. In all aspects
Author: Christopher P. Lapointe
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Book Description
Protein-RNA interactions are ubiquitous. They determine where, when, and how much protein is produced from an mRNA. Single RNA-binding proteins often bind to many RNAs. Multiple proteins can simultaneously bind to a single RNA molecule. The particular combination of proteins dictates the fate of that mRNA. Networks of protein-RNA interactions, referred to as 0́protein-RNA networks0́+, underlie fundamental cellular processes and have well-established connections to disease. The challenges now are to understand how they are formed, how they function, and how they are balanced in vivo. I employed an integrated approach of biochemistry, molecular biology, genomics, genetics, and bioinformatics to develop new strategies for the dissection of protein-RNA networks. I began with structure-function analyses of an RNA-modifying enzyme (Chapter 2), which enabled me to develop a method to identify protein-RNA interactions in vivo, which we termed 0́−RNA Tagging0́+ (Chapter 3). I used RNA Tagging to reveal that while proteins productively bind specific RNAs to control their function, they also 0́−sample0́+ RNAs by binding briefly to them without exerting a regulatory effect. I next integrated RNA Tagging with a different approach to identify RNAs bound by a particular protein called HITS-CLIP. I demonstrated that the two approaches are complementary, and together they defined a core set of mRNAs controlled by an RNA-binding protein (Chapter 4). These studies enabled collaborative multi-omic analyses, which revealed that Saccharomyces cerevisiae Puf3p is a key post-transcriptional repressor of mitochondrial biogenesis factors. In parallel, I employed RNA Tagging to dissect a protein-RNA network controlled by three related RNA-binding proteins 0́3 Puf3p, Puf4p, and Puf5p 0́3 in yeast (Chapter 5). The architecture of the network is controlled by competition among the proteins for mRNAs, which is balanced via an interplay of binding affinity and relative abundance. Together, my thesis research includes the development of a novel method and new strategies to analyze protein-RNA interactions that occur inside living cells. These approaches have several advantages over those previously described, provide new insights into how proteins regulate RNAs, and are widely applicable to protein-RNA networks throughout biology. They provide new opportunities to probe the nature and dynamics of protein-RNA networks in living cells.
Author: John Laver
Publisher:
ISBN:
Category :
Languages : en
Pages :
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Book Description
Post-transcriptional regulation of gene expression, through the control of mRNA splicing, polyadenylation, nuclear export, localization, translation, and stability, is essential for achieving appropriate temporal and spatial patterns of protein expression. This regulation is mediated by trans-acting factors, such as RNA-binding proteins (RBPs) and non-coding RNAs, which associate with specific mRNA targets through the recognition of sequence- or structure-based cis-elements present in the transcripts. The genomes of most organisms encode hundreds of RBPs, each of which likely associates with hundreds of mRNAs. Thus, a genome-wide view of the regulation being mediated by all trans-factors is essential for a complete understanding of post-transcriptional control. While post-transcriptional regulation is crucial in all biological systems, it has a particularly prominent role during early embryo development, as during this time there is no transcription from the zygotic genome of the embryo, and, thus, gene expression and development is controlled entirely post-transcriptionally. In this thesis, I describe my efforts towards obtaining a global understanding of post-transcriptional regulation in early Drosophila melanogaster embryos, through the development and use of synthetic antibodies as tools to identify, genome-wide, RBP-mRNA interactions. First, I demonstrated that synthetic antibodies generated against RBPs can be used as tools to identify RBP-associated mRNAs through immunoprecipitation-based approaches, or, conversely, to disrupt RBP-mRNA interactions. I then used synthetic antibodies to identify the entire complement of mRNAs associated with 3 developmentally-important RBPs: the double-stranded RBP Staufen, the TRIM-NHL protein Brain Tumor, and the PUF protein Pumilio. Computational analyses of these mRNAs revealed: (1) novel cis-elements likely mediating the mRNA-binding activity of Staufen and Brain Tumor; (2) that, unexpectedly, Brain Tumor and Pumilio function largely independently of each other in early embryos; and, (3) a novel role for Brain Tumor in promoting mRNA decay, which was demonstrated through a transcriptome-wide analysis of mRNA levels in brain tumor mutant embryos. To facilitate a truly genome-wide analysis of RBP-mRNA interactions, we developed a high-throughput pipeline for production of synthetic antibodies, and used this pipeline to generate 279 antibodies against 61 RBPs. In future this pipeline and the antibodies generated will allow for global studies of post-transcriptional regulation.
Author: Gene W. Yeo
Publisher: Springer
ISBN: 9783319804842
Category : Medical
Languages : en
Pages : 326
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Book Description
Ribonucleic acid (RNA) binding proteins currently number in the thousands and defects in their function are at the heart of diseases such as cancer and neurodegeneration. RNA binding proteins have become implicated in the intricate control of surprisingly diverse biological settings, such as circadian rhythm, stem cell self-renewal, oncogenesis and germ cell development. This book surveys a range of genome-wide and systems approaches to studying RNA binding proteins, the importance of RNA binding proteins in development, cancer and circadian rhythm.
Author: Nahum Sonenberg
Publisher: CSHL Press
ISBN: 9780879696184
Category : Gene expression
Languages : en
Pages : 1034
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Book Description
Since the 1996 publication of Translational Control, there has been fresh interest in protein synthesis and recognition of the key role of translation control mechanisms in regulating gene expression. This new monograph updates and expands the scope of the earlier book but it also takes a fresh look at the field. In a new format, the first eight chapters provide broad overviews, while each of the additional twenty-eight has a focus on a research topic of more specific interest. The result is a thoroughly up-to-date account of initiation, elongation, and termination of translation, control mechanisms in development in response to extracellular stimuli, and the effects on the translation machinery of virus infection and disease. This book is essential reading for students entering the field and an invaluable resource for investigators of gene expression and its control.
Author: Torben Heick Jensen
Publisher: Springer Science & Business Media
ISBN: 1441978410
Category : Medical
Languages : en
Pages : 161
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Book Description
The diversity of RNAs inside living cells is amazing. We have known of the more “classic” RNA species: mRNA, tRNA, rRNA, snRNA and snoRNA for some time now, but in a steady stream new types of molecules are being described as it is becoming clear that most of the genomic information of cells ends up in RNA. To deal with the enormous load of resulting RNA processing and degradation reactions, cells need adequate and efficient molecular machines. The RNA exosome is arising as a major facilitator to this effect. Structural and functional data gathered over the last decade have illustrated the biochemical importance of this multimeric complex and its many co-factors, revealing its enormous regulatory power. By gathering some of the most prominent researchers in the exosome field, it is the aim of this volume to introduce this fascinating protein complex as well as to give a timely and rich account of its many functions. The exosome was discovered more than a decade ago by Phil Mitchell and David Tollervey by its ability to trim the 3’end of yeast, S. cerevisiae, 5. 8S rRNA. In a historic account they laid out the events surrounding this identification and the subsequent birth of the research field. In the chapter by Kurt Januszyk and Christopher Lima the structural organization of eukaryotic exosomes and their evolutionary counterparts in bacteria and archaea are discussed in large part through presentation of structures.
Author: Krishnarao Appasani
Publisher: Cambridge University Press
ISBN: 9780521118552
Category : Science
Languages : en
Pages : 580
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Book Description
MicroRNAs (miRNAs) are RNA molecules, conserved by evolution, that regulate gene expressions and their recent discovery is revolutionising both basic biomedical research and drug discovery. Expression levels of MiRNAs have been found to vary between tissues and with developmental stages and hence evaluation of the global expression of miRNAs potentially provides opportunities to identify regulatory points for many different biological processes. This wide-ranging reference work, written by leading experts from both academia and industry, will be an invaluable resource for all those wishing to use miRNA techniques in their own research, from graduate students, post-docs and researchers in academia to those working in R&D in biotechnology and pharmaceutical companies who need to understand this emerging technology. From the discovery of miRNAs and their functions to their detection and role in disease biology, this volume uniquely integrates the basic science with industry application towards drug validation, diagnostic and therapeutic development. Forewords by: Sidney Altman, Yale University, Winner of the Nobel Prize in Chemistry, 1989 and Victor R. Ambros, Dartmouth Medical School, Co-discoverer of MicroRNAs
Author: Manfred Heinlein
Publisher: Humana
ISBN: 9781071607145
Category : Science
Languages : en
Pages : 490
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Book Description
This book provides a compendium of state-of-the-art methods for the labeling, detection, and purification of RNA and RNA-protein complexes and thereby constitutes an important toolbox for researchers interested in understanding the complex roles of RNA molecules in development, signaling, and disease. Beginning with a section on in situ detection of RNA molecules using FISH techniques, the volume continues with parts exploring in vivo imaging of RNA transport and localization, imaging and analysis of RNA uptake and transport between cells, identification and analysis of RNA-binding proteins, guide RNAs in genome editing, as well as other specific analytical techniques. Written for the highly successful Methods in Molecular Biology series, 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 practical, RNA Tagging: Methods and Protocols serves as a vital reference for researchers looking to further the increasingly important research in RNA biology.
Author: Imre Gaspar
Publisher: Humana Press
ISBN: 9781493972128
Category : Medical
Languages : en
Pages : 492
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Book Description
This volume introduces different concepts and methods of detecting RNA in biological material in a variety of model systems. The chapters in this book discuss methods that will answer numerous biological questions that arise in the study of RNAs. Some of the topics covered in this book are single mRNA molecule detection in embryos and neurons; detection of mRNA and associated molecules by ISH-IEM on frozen sections; optimizing molecular beacons for intracellular analysis of RNA; imaging translation dynamics of single mRNA molecules in live cells; preparation of high-throughput sequencing libraries; and capturing RNA binding proteins in embryos and in cell-culture. 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 comprehensive, RNA Detection: Methods and Protocols is a valuable resource for novel and experiences scientist in the expanding field of RNAs.
