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Bacterial Regulatory RNA pp 321–340Cite as

Techniques to Analyze sRNA Protein Cofactor Self-Assembly In Vitro

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1737))

Abstract

Post-transcriptional control of gene expression by small regulatory noncoding RNA (sRNA) needs protein accomplices to occur. Past research mainly focused on the RNA chaperone Hfq as cofactor. Nevertheless, recent studies indicated that other proteins might be involved in sRNA-based regulations. As some of these proteins have been shown to self-assemble, we describe in this chapter protocols to analyze the nano-assemblies formed. Precisely, we focus our analysis on Escherichia coli Hfq as a model, but the protocols presented here can be applied to analyze any polymer of proteins. This chapter thus provides a guideline to develop commonly used approaches to detect prokaryotic protein self-assembly, with a special focus on the detection of amyloidogenic polymers.

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Abbreviations

AFM:

Atomic force microscopy

CF:

Curve fitting

FTIR:

Fourier transform infrared spectroscopy

SAXS:

Small angle X-ray scattering

SRCD:

Synchrotron radiation circular dichroism

TEM:

Transmission electron microscopy

ThT:

Thioflavin T

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Acknowledgments

This work was supported by Université Paris Diderot, CNRS, CEA, and Synchrotron SOLEIL. We gratefully acknowledge help to MV from the French Embassy for their program for scientific and university cooperation. We are indebted to F. Gobeaux (CEA Saclay, Gif-sur-Yvette, France) and A. Deniset-Besseau (LCP, Université Paris-Sud, France) for many fruitful discussions. We thank Kimberly Stanek (University of Virginia) for her careful and critical reading of our manuscript.

Author information

Authors and Affiliations

  1. Synchrotron SOLEIL, L’Orme des Merisiers Saint Aubin, Gif-sur-Yvette, France

    David Partouche, Thomas Bizien, Christophe Sandt & Frank Wien

  2. Université Paris Diderot-Paris 7, Sorbonne Paris Cité, Paris, France

    David Partouche & Antoine Malabirade

  3. Instituto de Catálisis y Petroleoquímica, CSIC, Madrid, Spain

    Marisela Velez

  4. INSERM, U1196, Université Paris Sud, Université Paris-Saclay, Orsay, France

    Sylvain Trépout & Sergio Marco

  5. Institut Curie, PSL Research University, CNRS, UMR 91873348, Orsay, France

    Sylvain Trépout & Sergio Marco

  6. Department of Physics and Chemistry, University of Palermo, Palermo, Italy

    Valeria Militello

  7. Université Paris Diderot & LLB/CEA/CNRS, Gif-sur-Yvette, France

    Véronique Arluison

Authors
  1. David Partouche
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  2. Antoine Malabirade
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  3. Thomas Bizien
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  4. Marisela Velez
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  5. Sylvain Trépout
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  6. Sergio Marco
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  7. Valeria Militello
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  8. Christophe Sandt
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  9. Frank Wien
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  10. Véronique Arluison
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Corresponding author

Correspondence to Véronique Arluison .

Editor information

Editors and Affiliations

  1. Université Paris Diderot & LLB/CEA/CNRS, Gif-sur-Yvette, France

    Véronique Arluison

  2. Laboratorio de Bioquímica, Microbiología e Interacciones, Biológicas en el Suelo, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes-CONICET, Bernal, Argentina

    Claudio Valverde

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Partouche, D. et al. (2018). Techniques to Analyze sRNA Protein Cofactor Self-Assembly In Vitro. In: Arluison, V., Valverde, C. (eds) Bacterial Regulatory RNA. Methods in Molecular Biology, vol 1737. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7634-8_18

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  • DOI: https://doi.org/10.1007/978-1-4939-7634-8_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7633-1

  • Online ISBN: 978-1-4939-7634-8

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