Influence of Splicing Factors on Recognition and Utilization of Mammalian Pre-mRNA 3' Processing Sites PDF Download
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Author: Jonathan Joseph Cherry
Publisher:
ISBN:
Category :
Languages : en
Pages : 362
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Author: Jonathan Joseph Cherry
Publisher:
ISBN:
Category :
Languages : en
Pages : 362
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Author: Karli Lipinski
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Book Description
Pre-mRNA splicing is a fundamental process governing gene expression in eukaryotes and is orchestrated by the spliceosome, a dynamic ribonucleoprotein complex. Spliceosomes catalyze splicing in two steps and are composed of both small nuclear RNAs (snRNAs; U1, U2, U4, U5, U6) and numerous proteins. Dynamic interactions between snRNAs, proteins, and the pre-mRNA substrate occur during splicing which are central to regulating splice site recognition, catalysis, and fidelity. This thesis examines multiple facets of RNA-protein interactions within spliceosome complexes beginning with snRNA biogenesis, through formation of the spliceosome active site, and during the second step of chemistry, exon ligation. Functional mechanisms into splicing, such as selection of intron recognition sites, are examined. Function of the main catalytic component of the spliceosome, the U6 snRNA, is first explored. Transcription and post-transcriptional processing of U6 is unique among snRNAs and therefore may be required to generate a functional U6. Transcription by RNAP II instead of RNAP III produces a functional U6 molecule. Defects in stability, likely a result of incorrect post-transcriptional processing and binding of stabilizing proteins, result in changes to in vivo distributions of spliceosome sub particles called snRNPs. Transcription of U6 by RNAP II is useful for the incorporation of genetic tags for endogenous fluorescent labeling or purification. Additionally, new applications of endogenous fluorescent labeling techniques within the U4 snRNA are presented, paving the way for single molecule studies of snRNA dynamics and Brr2 helicase function during activation. Mango and MS2 tags are well tolerated in the yeast U4 snRNA and with several of the tagged U4 constructs also minimally impacting splicing activities. Future single molecule experiments will examine the timing of the U4 snRNA release from the spliceosome compared to the release of Prp3, a protein associated with U6 and U4 snRNAs. Finally, validation of a proposed novel splicing factor, Fyv6, and study of its influence on 3' SS represent major contributions to the field of splicing. Utilizing a new high resolution P complex structure of the spliceosome containing Fyv6 solved by Max Wilkinson, I examined multiple contacts of Fyv6 with other splicing factors, notably Prp22. Genetic studies in yeast show that interactions with the protein Syf1 and Prp22 are important for Fyv6 function. The absence of Fyv6 from spliceosomes results in transcriptome-wide splicing defects, largely due to changes in 3' SS usage. From these studies, Fyv6 can be added to the list of splicing factors that impact the second step of splicing and affect fidelity of 3' SS selection.
Author: Adrian Krainer
Publisher: IRL Press
ISBN:
Category : Language Arts & Disciplines
Languages : en
Pages : 408
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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: Klemens J. Hertel
Publisher: Humana
ISBN: 9781627039796
Category : Medical
Languages : en
Pages : 0
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Book Description
Providing a guide to classical experimental approaches to decipher splicing mechanisms and experimental strategies that rely on novel multi-disciplinary approaches, Spliceosomal Pre-mRNA Splicing: Methods and Protocols describes the theory of alternative pre-mRNA splicing in seven introductory chapters and then introduces protocols and their theoretical background relevant for a variety of experimental research. These protocol chapters cover basic methods to detect splicing events, analyses of alternative pre-mRNA splicing in vitro and in vivo manipulation of splicing events and high-throughput and bioinformatic analyses of alternative splicing. 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 protocols and tips on troubleshooting and avoiding known pitfalls. Comprehensive and practical, Spliceosomal Pre-mRNA Splicing: Methods and Protocols will aid newcomers and seasoned molecular biologists in understanding the fascinating world of alternative splicing with the ultimate goal of paving the way for many new discoveries to come.
Author: Philippe Jeanteur
Publisher: Springer Science & Business Media
ISBN: 3540344497
Category : Science
Languages : en
Pages : 265
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Book Description
Splicing of primary RNA transcript is a quasi-systematic step of gene expression in higher organisms. This is the first book to highlight the medical implications, i.e. diseases, caused by alternative splicing. Alternative splicing not only vastly increases protein diversity but also offers numerous opportunities for aberrant splicing events with pathological consequences. The book also outlines possible targets for therapy.
Author: Rosanna Asselta
Publisher: Frontiers Media SA
ISBN: 2889662357
Category : Science
Languages : en
Pages : 197
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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.
Author: B. D. Hames
Publisher: Oxford University Press, USA
ISBN:
Category : Music
Languages : en
Pages : 238
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Book Description
This book gives a co-ordinated review of our present knowledge of eukaryotic RNA synthesis.
Author: Philippe Jeanteur
Publisher:
ISBN: 9783662097298
Category :
Languages : en
Pages : 266
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Author:
Publisher:
ISBN: 9780815332183
Category : Cells
Languages : en
Pages : 0
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Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 1844
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