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Riboswitches
Methods and Protocols
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Main description:

The transfer of hereditary information from genes to proteins is one of the essential pr- esses in all living organisms on our planet. Some genes are expressed without modu- tion throughout the life of a cell, while many others require various degrees of control to precisely balance cellular metabolism with environmental conditions. For many years, researchers attributed this regulatory function to protein molecules, which can direct gene expression at multiple levels, in response to various input signals, and with different degrees of selectivity. Even when the control of gene expression was achieved via direct interactions between proteins and mRNAs, the active role was routinely assigned to p- teins, while RNAs were considered merely as recipient molecules. The discovery of RNA interference and multiple bacterial regulatory RNAs caused a shift from the perception of proteins as the predominant regulators of gene expression to the acknowledgement of the importance of RNAs in many regulatory circuits. Such a viewpoint received strong support several years ago after the discovery of riboswitches and related RNA sensors - mRNA regions capable of alternating their conformations in response to the presence of cellular metabolites and other physical or chemical cues. These classes of RNA pass on cellular and environmental information directly to transcription or translation machinery without the assistance of proteins. The riboswitches are commonly defined as evolutionarily conserved mRNA regions capable of specific binding to metabolite molecules, and, as a result, adopting a particular RNA conformation that modulates gene expression.


Contents:

1. Predicting Riboswitch Regulation on a Genomic Scale
Jeffrey E. Barrick

2. Enzymatic Ligation Strategies for the Preparation of Purine Riboswitches with Site-Specific Chemical Modifications
Renate Rieder, Claudia Hoebartner, and Ronald Micura

3. Application of Fluorescent Measurements for Characterization of Riboswitch-Ligand Interactions
Benoit Heppell, Jerome Mulhbacher, J. Carlos Penedo, and Daniel A. Lafontaine

4. Transcriptional Approaches to Riboswitch Studies
Alexander Mironov, Vitaly Epshtein, and Evgeny Nudler

5. Kinetics of Riboswitch Regulation Studied By In Vitro Transcription
J. Kenneth Wickiser

6. Molecular Basis of RNA-Mediated Gene Regulation on the Adenine Riboswitch by Single-Molecule Approaches
Jean-Francois Lemay, J. Carlos Penedo, Jerome Mulhbacher, and Daniel A. Lafontaine

7. Methods for Analysis of Ligand-Induced RNA Conformational Changes
Chad A. Brautigam, Catherine A. Wakeman, and Wade C. Winkler

8. Monitoring RNA-Ligand Interactions Using Isothermal Titration Calorimetry
Sunny D. Gilbert and Robert T. Batey

9. Preparation and Crystallization of Riboswitch-Ligand Complexes
Olga Pikovskaya, Artem A. Serganov, Anna Polonskaia, Alexander Serganov, and Dinshaw J. Patel

10. Crystallization of the glmS Ribozyme-Riboswitch
Daniel J. Klein and Adrian R. Ferre-D'Amare

11. Riboswitch Conformations Revealed by Small-Angle X-Ray Scattering
Jan Lipfert, Daniel Herschlag, and Sebastian Doniach

12. Time-Resolved NMR Spectroscopy: Ligand-Induced Refolding of Riboswitches
Janina Buck, Boris Furtig, Jonas Noeske, Jens Woehnert, and Harald Schwalbe

13. Analysis of the RNA Backbone: Structural Analysis of Riboswitches by In-Line Probing and Selective 2'-Hydroxyl Acylation and PrimerExtension (SHAPE)
Catherine A. Wakeman and Wade C. Winkler

14. Identification of Metabolite-Riboswitch Interactions Using Nucleotide Analog Interference Mapping and Suppression
Juliane K. Soukup and Garrett A. Soukup

15. RNA-Dependent RNA Switches in Bacteria
Tina M. Henkin

16. Probing mRNA Structure and sRNA-mRNA Interactions in Bacteria Using Enzymes and Lead(II)
Clement Chevalier, Thomas Geissmann, Anne-Catherine Helfer, and Pascale Romby

17. Structural Probing of RNA Thermosensors
Claude Chiaruttini, Frederic Allemand, and Mathias Springer

18. Ribosomal Initiation Complexes Probed by Toeprinting and Effect of trans-Acting Translational Regulators in Bacteria
Pierre Fechter, Clement Chevalier, Gulnara Yusupova, Marat Yusupov, Pascale Romby, and Stefano Marzi

19. Isolation and Characterization of the Heat Shock RNA 1 (HSR1)
Ilya Shamovsky and Evgeny Nudler

20. Analysis of tRNA-Directed Transcription Antitermination in the T Box System in vivo
Tina M. Henkin

21. In vitro Selection of Conformational Probes for Riboswitches
Gunter Mayer and Michael Famulok

22. A Green Fluorescent Protein (GFP) Based Plasmid System to Study Post-Transcriptional Control of Gene Expression in vivo
Johannes H. Urban and Joerg Vogel

23. High-Throughput Screens to Discover Synthetic Riboswitches
Sean A. Lynch, Shana Topp, and Justin P. Gallivan

24. A Mammalian Cell-Based Assay for Screening Inhibitors of RNA Cleavage
Laising Yen, Brent R. Stockwell, and Richard C. Mulligan
25. In vitro Selection of glmS Ribozymes
Kristian H. Link and Ronald R. Breaker


PRODUCT DETAILS

ISBN-13: 9781617379475
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 377
Weight: 934g
Availability: Available
Subcategories: Biochemistry, Genetics
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