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Riboswitches

Methods and Protocols

  • A comprehensive and timely reference volume for basic and applied research on regulatory RNAs
  • Covers a wide range of assay formats and systems from the computerized prediction of regulatory RNA elements to their detailed structure-function characterization
  • Focuses on both natural and synthetic RNA molecules
  • Provides step-by-step protocols of rigorously tested classical and cutting-edge in vivo and in vitro methods
  • Presents techniques easily adaptable for experiments with any RNA species
  • Includes supplementary material: sn.pub/extras

Part of the book series: Methods in Molecular Biology (MIMB, volume 540)

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Table of contents (25 protocols)

  1. Front Matter

    Pages i-xi
  2. Application of Fluorescent Measurements for Characterization of Riboswitch–Ligand Interactions

    • Benoit Heppell, Jérôme Mulhbacher, J. Carlos Penedo, Daniel A. Lafontaine
    Pages 25-37
  3. Transcriptional Approaches to Riboswitch Studies

    • Alexander Mironov, Vitaly Epshtein, Evgeny Nudler
    Pages 39-51
  4. Molecular Basis of RNA-Mediated Gene Regulation on the Adenine Riboswitch by Single-Molecule Approaches

    • Jean-François Lemay, J. Carlos Penedo, Jérôme Mulhbacher, Daniel A. Lafontaine
    Pages 65-76
  5. Methods for Analysis of Ligand-Induced RNA Conformational Changes

    • Chad A. Brautigam, Catherine A. Wakeman, Wade C. Winkler
    Pages 77-95
  6. Preparation and Crystallization of Riboswitch–Ligand Complexes

    • Olga Pikovskaya, Artem A. Serganov, Ann Polonskaia, Alexander Serganov, Dinshaw J. Patel
    Pages 115-128
  7. Crystallization of the glmS Ribozyme-Riboswitch

    • Daniel J. Klein, Adrian R. Ferré-D’Amaré
    Pages 129-139
  8. Riboswitch Conformations Revealed by Small-Angle X-Ray Scattering

    • Jan Lipfert, Daniel Herschlag, Sebastian Doniach
    Pages 141-159
  9. Time-Resolved NMR Spectroscopy: Ligand-Induced Refolding of Riboswitches

    • Janina Buck, Boris Fürtig, Jonas Noeske, Jens Wöhnert, Harald Schwalbe
    Pages 161-171
  10. RNA-Dependent RNA Switches in Bacteria

    • Tina M. Henkin
    Pages 207-214
  11. Probing mRNA Structure and sRNA–mRNA Interactions in Bacteria Using Enzymes and Lead(II)

    • Clément Chevalier, Thomas Geissmann, Anne-Catherine Helfer, Pascale Romby
    Pages 215-232
  12. Structural Probing of RNA Thermosensors

    • Claude Chiaruttini, Frédéric Allem, Mathias Springer
    Pages 233-245
  13. Ribosomal Initiation Complexes Probed by Toeprinting and Effect of trans-Acting Translational Regulators in Bacteria

    • Pierre Fechter, Clément Chevalier, Gulnara Yusupova, Marat Yusupov, Pascale Romby, Stefano Marzi
    Pages 247-263
  14. Isolation and Characterization of the Heat Shock RNA 1

    • Ilya Shamovsky, Evgeny Nudler
    Pages 265-279

About this book

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.

Reviews

From the reviews:

"Riboswitches are among the latest additions to the growing field of RNA-based gene expression control systems found in both prokaryotic and eukaryotic cells. … useful in various aspects of work on riboswitches and other regulatory RNAs. … It also refers to pertinent and up-to-date literature. … The reviewed book can be strongly recommended to RNA researchers with different levels of expertise, as well as to those who learn or teach in this field." (Joanna Ciesla, Acta Biochimica Polonica, Vol. 56 (3), 2009)

“This book is a good collection of ‘Methods and Protocols’ for studying … riboswitches as well as some other RNAs involved in gene expression. … Readers are introduced to a broad range of RNA methodologies from basic to applied in this book. I … recommend the book for researchers who work in the area of riboswitches, but also for other scientists who are involved in RNA-relevant studies.” (Atsushi Ogawa, ChemBioChem, Vol. 10, 2009)

Bibliographic Information

  • Book Title: Riboswitches

  • Book Subtitle: Methods and Protocols

  • Editors: Alexander Serganov

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-59745-558-9

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 2009

  • Hardcover ISBN: 978-1-934115-88-6Published: 17 April 2009

  • Softcover ISBN: 978-1-61737-947-5Published: 19 November 2010

  • eBook ISBN: 978-1-59745-558-9Published: 21 April 2009

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 366

  • Number of Illustrations: 65 b/w illustrations

  • Topics: Biochemistry, general, Nucleic Acid Chemistry, Gene Expression

Buy it now

Buying options

eBook USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Other ways to access