Isolation and Characterization of Splicing Regulatory Elements from the C-src Pre-messenger RNA PDF Download
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Author: Edward Frank Modafferi
Publisher:
ISBN:
Category :
Languages : en
Pages : 238
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Author: Edward Frank Modafferi
Publisher:
ISBN:
Category :
Languages : en
Pages : 238
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Author: Scott Adamson
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Alternative splicing is a key process in higher eukaryotes in which different introns in pre-mRNA are removed to generate diversity in mature RNA products. Changes in alternative splicing are important under normal conditions for expanding the proteome and regulating RNA species, however splicing dysregulation can underlie monogenic and polygenic diseases. Understanding the connection between genetic variants and phenotypes is a major goal of personalized medicine and is key to understanding the underlying biological processes. Here I develop a novel highthroughput splicing reporter assay to elucidate the impact of genetic variants on pre-mRNA splicing efficiency called variant exon sequencing (Vex-seq). The utility of Vex-seq is first demonstrated in the context of thousands of variants found in the human population across 110 exons, then it is applied to understand the sequence determinants of splicing regulatory elements. These splicing regulatory elements were identified by examining ENCODE data which monitors RNA-protein interactions, and changes in splicing upon RNA binding protein (RBP) knockdown. Vex-seq is used to identify sequence features associated with RBP-specific splicing regulation and contribute to our understanding of a key, understudied mechanistic link between sequence variation and splicing regulation.
Author: Hosung Min
Publisher:
ISBN:
Category :
Languages : en
Pages : 350
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Author: Stefan Stamm
Publisher: John Wiley & Sons
ISBN: 3527647988
Category : Science
Languages : en
Pages : 660
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Book Description
This book was written for graduate and medical students, as well as clinicians and postdoctoral researchers. It describes the theory of alternative pre-mRNA splicing in twelve introductory chapters and then introduces protocols and their theoretical background relevant for experimental research. These 43 practical chapters cover: Basic methods, Detection of splicing events, Analysis of alternative pre-mRNA splicing in vitro and in vivo, Manipulation of splicing events, and Bioinformatic analysis of alternative splicing. A theoretical introduction and practical guide for molecular biologists, geneticists,clinicians and every researcher interested in alternative splicing. Website: www.wiley-vch.de/home/splicing
Author: Adrian Krainer
Publisher: IRL Press
ISBN:
Category : Language Arts & Disciplines
Languages : en
Pages : 408
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Book Description
This volume focuses on the major aspects of post-transcriptional mRNA processing in the nucleus of eukaryotic cells. Each of the described mRNA reactions is required for proper gene expression and can also serve as a control point for regulating the expression of many genes, for example duringembryonic development or in different cell types. The different chapters review the assembly of newly synthesized nuclear mRNA transcripts into hnRNP particles and catalytically active spliceosomes; the structure and mechanism of action of small nuclear ribonucleoprotein particles and proteinfactors that catalyse pre-mRNA splicing in mammalian cells and in yeast; the regulation of gene expression and generation of protein isoform diversity by alternative splicing; the mechanisms of 3' end cleavage and polyadenylation; the architecture of the cell nucleus in relation to these processesand to the localization of the relevant substrates and factors; the diverse mechanisms of RNA processing by ribozymes and their potential relevance for nuclear mRNA processing; the mechanism of spliced-leader addition by trans-splicing in nematodes and trypanosomes; and the process ofinsertion/deletion mRNA editing in kinetoplasmid protozoa. In each chapter, leading researchers have provided detailed, critical reviews of the history, experimental approaches, major advances, current ideas and models, as well as future directions, for each of these active areas of research.
Author: Angela Norie Brooks
Publisher:
ISBN:
Category :
Languages : en
Pages : 236
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A majority of metazoan genes contain introns in their primary transcripts (pre-mRNA) that require removal by the spliceosome--a cellular complex composed of protein and RNA. Upon removal of introns from the primary transcript, the remaining exonic portion of the transcript is spliced together. It is essential to remove the correct intronic portion of a primary transcript in order to produce the desired product, typically a protein-coding mRNA. Pre-mRNAs are alternatively spliced when different intron boundaries are used by the spliceosome, thus creating different mRNA products. Alternative splicing allows for an additional step of gene regulation by producing transcript isoforms that can be differentially processed in a particular tissue or developmental time point. Alternative splicing is primarily regulated by RNA binding proteins that bind to pre-mRNA and act to recruit or inhibit the spliceosome at specific splice sites. A central aim of this work is to gain a better understanding of splicing regulation by the identification and characterization of protein regulators of splicing and cis-acting splicing regulatory sequences in the model organism, Drosophila melanogaster. To identify splicing regulatory elements, many previous studies in vertebrate genomes have used computational methods. In collaboration with Anna I. Podgornaia, I applied such an approach to predict splicing regulatory elements in Drosophila melanogaster and compared them with elements found in vertebrates. I identified 330 putative splicing enhancer sequences enriched near weak 5' and 3' splice sites of constitutively spliced introns. I found that a significant proportion (58%) of D. melanogaster enhancers were previously reported as splicing enhancers in vertebrates. To provide additional evidence for the function of the intronic splicing enhancers (ISEs), I identified intronic hexamers significantly enriched within sequences phylogenetically conserved among 15 insect species. This analysis uncovered 73 putative ISEs that are also enriched in conserved regions of the D. melanogaster genome. The functions of nine enhancer sequences were verified in a heterologous splicing reporter by Julie L. Aspden, demonstrating that these sequences are sufficient to enhance splicing in vivo. Taken together, these data identify a set of predicted positive-acting splicing regulatory motifs in the Drosophila genome and highlight those regulatory sequences that are present in distant metazoan genomes. To identify and characterize splicing regulators, collaborators and I have combined RNAi and RNA-Seq to identify exons that are regulated by 58 known or putative splicing regulators. To identify and quantify alternative splicing events from RNA-Seq data, I developed the JuncBASE (Junction Based Analysis of Splicing Events) software package. For a pilot study, I identified 404 splicing events significantly affected upon depletion of pasilla. Preliminary analysis showed 879 splicing events affected by at least one of the 57 other proteins. The sequence regions upstream and within Pasilla-repressed exons and downstream of Pasilla-activated exons are enriched for YCAY repeats, which is consistent with the location of these motifs near regulated exons of the mammalian ortholog, Nova. Thus, the RNA regulatory map of Pasilla and Nova is highly conserved between insects and mammals despite the fact that the pre-mRNAs that are regulated by Pasilla and Nova are almost entirely non-overlapping. This observation strongly suggests that the regulatory codes of individual RNA binding proteins are nearly immutable, yet the regulatory modules controlled by these proteins are highly evolvable. I also present RNA regulatory maps for the four hnRNP proteins: hrp36, hrp38, hrp40, and hrp48. Lastly, I examine splicing regulation throughout the life cycle of D. melanogaster. Using transcriptome data from 30 developmental time points produced by collaborators from the modENCODE Consortium, I identified a total of 23,859 alternative splicing events in Drosophila, taking into account all transcript information from D. melanogaster annotations, short sequenced reads (Illumina RNA-Seq), sequenced cDNA, long RNA-Seq reads (454 RNA-Seq) from adult flies, and short read sequences of rRNA-depleted RNA from embryonic time points. I observed that 60.7% of intron-containing genes in D. melanogaster are alternatively spliced. Using only the Illumina RNA-Seq reads throughout development, 21,216 splicing events were expressed and 13,951 events were differentially spliced in at least one time point. I also observed exons with similar patterns of splicing changes throughout development as well as sex-biased alternative splicing. Integrating information from our pasilla study, I also observed correlations of pasilla gene expression with alternative splicing changes of its target exons throughout development.
Author:
Publisher:
ISBN:
Category : Dissertations, Academic
Languages : en
Pages : 794
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Author: Philippe Jeanteur
Publisher:
ISBN: 9783662097298
Category :
Languages : en
Pages : 266
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Author: Winslow R. Briggs
Publisher: John Wiley & Sons
ISBN: 3527604855
Category : Science
Languages : en
Pages : 496
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Book Description
This first complete resource on photosensory receptors from bacteria, plants and animals compiles the data on all known classes of photoreceptors, creating a must-have reference for students and researchers for many years to come. Among the editors are the current and a former president of the American Society for Photobiology.
Author: Ezequiel Petrillo
Publisher: Frontiers Media SA
ISBN: 2889639746
Category : Nature
Languages : en
Pages : 175
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Book Description
This eBook is a collection of articles from a Frontiers Research Topic. Frontiers Research Topics are very popular trademarks of the Frontiers Journals Series: they are collections of at least ten articles, all centered on a particular subject. With their unique mix of varied contributions from Original Research to Review Articles, Frontiers Research Topics unify the most influential researchers, the latest key findings and historical advances in a hot research area! Find out more on how to host your own Frontiers Research Topic or contribute to one as an author by contacting the Frontiers Editorial Office: frontiersin.org/about/contact.
