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Biochemical Studies of the Eukaryotic RNA Exosome

Author : Matthew Alan Bratkowski
Publisher :
Page : 209 pages
File Size : 12,60 MB
Release : 2012
Category :
ISBN :

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The RNA exosome is a multi-subunit protein complex involved in RNA maturation, surveillance, and turnover of cellular RNA. It is composed of a nine subunit core that contains an RNA binding central channel and associates with the additional subunit Rrp44. Rrp44 contains a 3' to 5' processive exoribonuclease activity and an endoribonuclease activity in its N-terminal PIN domain. The exosome is present in both the nucleus and cytoplasm. In the nucleus, the exosome associates with a second nuclease, Rrp6, which is a distributive 3' to 5' exoribonuclease. Although a wealth of studies have helped to characterize the exosome, it is still not entirely clear how the exosome core functions in concert with these two nucleases to degrade RNA. In this dissertation, I present structural and biochemical studies of both the Rrp44 and Rrp6 bound exosome. In particular, I have attempted to crystallize a sub-complex of Rrp44 bound to the exosome, and present data that suggest that structured RNA can access Rrp44 without being first channeled through the exosome. Despite this conclusion, the exosome core still appears to down-regulate both the exonuclease and endonuclease activities of Rrp44, possibly by causing the protein to assume a conformation that prohibits it from promiscuous RNA degradation. Work on the Rrp6 containing exosme was conducted in attempts of obtaining an EM reconstruction of this complex. Crystallization of an RNA and Rrp6 binding exosome associated iii protein, Rrp47, was also attempted alone and in complex with Rrp6. Overall, the results that I present here will help to further characterize the function of the exosome in RNA mediated degradation. iv.

The Eukaryotic RNA Exosome

Author : John LaCava
Publisher : Springer Nature
Page : 514 pages
File Size : 11,96 MB
Release : 2019-11-25
Category : Medical
ISBN : 1493998226

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This volume provides a cross-section of RNA exosome research protocols, applied to a diversity of model organisms. Chapters guide readers through methods that e.g. delineate eukaryotic exosomes’ origins in prokaryotes, probe its RNA substrates, adapter complexes and macromolecular interaction of networks, and establish critical structural-function relationships. 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 cutting-edge, The Eukaryotic RNA Exosome: Methods and Protocols aims to ensure successful results in the further study of this vital field.

RNA Exosome

Author : Torben Heick Jensen
Publisher : Springer Science & Business Media
Page : 161 pages
File Size : 48,94 MB
Release : 2011-06-29
Category : Medical
ISBN : 1441978410

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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.

Eukaryotic RNases and their Partners in RNA Degradation and Biogenesis

Author :
Publisher : Academic Press
Page : 295 pages
File Size : 14,67 MB
Release : 2012-12-31
Category : Science
ISBN : 0124047459

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This special issue of The Enzymes is targeted towards researchers in biochemistry, molecular and cell biology, pharmacology, and cancer. This volume discusses Eukaryotic RNases and their partners in RNA degradation and biogenesis. Contributions from leading authorities Informs and updates on all the latest developments in the field

RNA Turnover in Eukaryotes: Nucleases, Pathways and Analysis of mRNA Decay

Author : Lynne E. Maquat
Publisher : Academic Press
Page : 661 pages
File Size : 22,13 MB
Release : 2009-01-30
Category : Science
ISBN : 0080922074

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Specific complexes of protein and RNA carry out many essential biological functions, including RNA processing, RNA turnover, RNA folding, as well as the translation of genetic information from mRNA into protein sequences. Messenger RNA (mRNA) decay is now emerging as an important control point and a major contributor to gene expression. Continuing identification of the protein factors and cofactors, and mRNA instability elements responsible for mRNA decay allow researchers to build a comprehensive picture of the highly orchestrated processes involved in mRNA decay and its regulation. Covers the nonsense-mediated mRNA decay (NMD) or mRNA surveillance pathway Expert researchers introduce the most advanced technologies and techniques to identify mRNA processing, transport, localization and turnover, which are central to the process of gene expression Offers step-by-step lab instructions, including necessary equipment and reagents

Eukaryotic RNases and their Partners in RNA Degradation and Biogenesis

Author :
Publisher : Academic Press
Page : 0 pages
File Size : 19,55 MB
Release : 2012-11-27
Category : Science
ISBN : 9780124047419

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This special volume of The Enzymes is targeted towards researchers in biochemistry, molecular and cell biology, pharmacology, and cancer. This thematic volume discusses Eukaryotic RNases and their partners in RNA degradation and biogenesis.

RNA Metabolism and Gene Expression in Archaea

Author : Béatrice Clouet-d'Orval
Publisher : Springer
Page : 276 pages
File Size : 13,61 MB
Release : 2017-10-28
Category : Science
ISBN : 331965795X

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This book focuses on the regulation of transcription and translation in Archaea and arising insights into the evolution of RNA processing pathways. From synthesis to degradation and the implications of gene expression, it presents the current state of knowledge on archaeal RNA biology in 13 chapters. Topics covered include the modification and maturation of RNAs, the function of small non-coding RNAs and the CRISPR-Cas defense system. While Archaea have long been considered exotic microbial extremophiles, they are now increasingly being recognized as important model microorganisms for the study of molecular mechanisms conserved across the three domains of life, and with regard to the relevance of similarities and differences to eukaryotes and bacteria. This unique book offers a valuable resource for all readers interested in the regulation of gene expression in Archaea and RNA metabolism in general.

Fungal Extracellular Vesicles

Author : Marcio Rodrigues
Publisher : Springer Nature
Page : 193 pages
File Size : 39,56 MB
Release : 2022-01-01
Category : Science
ISBN : 3030833917

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This book provides an in-depth overview on the manifold functions of fungal extracellular vesicles (EV) which span from cell-to-cell communication, pathogenicity and stimulation of host’s immunity to export of hundreds of biomolecules. The book summarizes the present knowledge on the impact of extracellular vesicles on fungal biology. Extracellular vesicles participate in fundamental biological processes in all living cells but only during the last 15 years the production and functions of EVs were identified and studied in fungal species too. Up to date more than 50 independent studies have shown that extracellular vesicles are produced by at least 20 fungal species. The book addresses researchers and advanced students in Microbiology, Mycology and Biotechnology.

Untranslated Gene Regions and Other Non-coding Elements

Author : Lucy W. Barrett
Publisher : Springer Science & Business Media
Page : 63 pages
File Size : 22,91 MB
Release : 2013-06-26
Category : Science
ISBN : 3034806795

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There is now compelling evidence that the complexity of higher organisms correlates with the relative amount of non-coding RNA rather than the number of protein-coding genes. Previously dismissed as “junk DNA”, it is the non-coding regions of the genome that are responsible for regulation, facilitating complex temporal and spatial gene expression through the combinatorial effect of numerous mechanisms and interactions working together to fine-tune gene expression. The major regions involved in regulation of a particular gene are the 5’ and 3’ untranslated regions and introns. In addition, pervasive transcription of complex genomes produces a variety of non-coding transcripts that interact with these regions and contribute to regulation. This book discusses recent insights into the regulatory roles of the untranslated gene regions and non-coding RNAs in the control of complex gene expression, as well as the implications of this in terms of organism complexity and evolution.​