A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions

Pierre Olivier Vidalain; Yves Jacob; Marne C. Hagemeijer; Louis M. Jones; Grégory Neveu; Jean Pierre Roussarie; Peter J M Rottier; Frédéric Tangy; Cornelis A M De Haan

doi:10.1007/978-1-4939-2438-7_18

A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions

Pierre Olivier Vidalain^*
, Yves Jacob
, Marne C. Hagemeijer
, Louis M. Jones
, Grégory Neveu
, Jean Pierre Roussarie
, Peter J M Rottier
, Frédéric Tangy
, Cornelis A M De Haan

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

Abstract

Over the last 2 decades, yeast two-hybrid became an invaluable technique to decipher protein-protein interaction networks. In the field of virology, it has proven instrumental to identify virus-host interactions that are involved in viral embezzlement of cellular functions and inhibition of immune mechanisms. Here, we present a yeast two-hybrid protocol that has been used in our laboratory since 2006 to search for cellular partners of more than 300 viral proteins. Our aim was to develop a robust and straightforward pipeline, which minimizes false-positive interactions with a decent coverage of target cDNA libraries, and only requires a minimum of equipment. We also discuss reasons that motivated our technical choices and compromises that had to be made. This protocol has been used to screen most non-structural proteins of murine hepatitis virus (MHV), a member of betacoronavirus genus, against a mouse brain cDNA library. Typical results were obtained and are presented in this report.

Original language	English
Title of host publication	Coronaviruses: Methods and Protocols
Editors	Helena Jane Maier, Erica Bickerton, Paul Britton
Publisher	Springer New York
Pages	213-229
Number of pages	17
Volume	V
ISBN (Electronic)	9781493924387
ISBN (Print)	9781493924370
DOIs	https://doi.org/10.1007/978-1-4939-2438-7_18
Publication status	Published - 26 Feb 2015

Publication series

Name	Methods in Molecular Biology
Volume	1282
ISSN (Print)	1064-3745

Keywords

Host-pathogen interactions
Interactomics
Murine hepatitis virus
Proteomics
Yeast two-hybrid

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10.1007/978-1-4939-2438-7_18

Cite this

Vidalain, P. O., Jacob, Y., Hagemeijer, M. C., Jones, L. M., Neveu, G., Roussarie, J. P., Rottier, P. J. M., Tangy, F., & De Haan, C. A. M. (2015). A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions. In H. J. Maier, E. Bickerton, & P. Britton (Eds.), Coronaviruses: Methods and Protocols (Vol. V, pp. 213-229). (Methods in Molecular Biology; Vol. 1282). Springer New York. https://doi.org/10.1007/978-1-4939-2438-7_18

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abstract = "Over the last 2 decades, yeast two-hybrid became an invaluable technique to decipher protein-protein interaction networks. In the field of virology, it has proven instrumental to identify virus-host interactions that are involved in viral embezzlement of cellular functions and inhibition of immune mechanisms. Here, we present a yeast two-hybrid protocol that has been used in our laboratory since 2006 to search for cellular partners of more than 300 viral proteins. Our aim was to develop a robust and straightforward pipeline, which minimizes false-positive interactions with a decent coverage of target cDNA libraries, and only requires a minimum of equipment. We also discuss reasons that motivated our technical choices and compromises that had to be made. This protocol has been used to screen most non-structural proteins of murine hepatitis virus (MHV), a member of betacoronavirus genus, against a mouse brain cDNA library. Typical results were obtained and are presented in this report.",

keywords = "Host-pathogen interactions, Interactomics, Murine hepatitis virus, Proteomics, Yeast two-hybrid",

author = "Vidalain, \{Pierre Olivier\} and Yves Jacob and Hagemeijer, \{Marne C.\} and Jones, \{Louis M.\} and Gr{\'e}gory Neveu and Roussarie, \{Jean Pierre\} and Rottier, \{Peter J M\} and Fr{\'e}d{\'e}ric Tangy and \{De Haan\}, \{Cornelis A M\}",

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doi = "10.1007/978-1-4939-2438-7\_18",

language = "English",

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Vidalain, PO, Jacob, Y, Hagemeijer, MC, Jones, LM, Neveu, G, Roussarie, JP, Rottier, PJM, Tangy, F & De Haan, CAM 2015, A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions. in HJ Maier, E Bickerton & P Britton (eds), Coronaviruses: Methods and Protocols. vol. V, Methods in Molecular Biology, vol. 1282, Springer New York, pp. 213-229. https://doi.org/10.1007/978-1-4939-2438-7_18

A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions. / Vidalain, Pierre Olivier; Jacob, Yves; Hagemeijer, Marne C. et al.
Coronaviruses: Methods and Protocols. ed. / Helena Jane Maier; Erica Bickerton; Paul Britton. Vol. V Springer New York, 2015. p. 213-229 (Methods in Molecular Biology; Vol. 1282).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

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AU - Jacob, Yves

AU - Hagemeijer, Marne C.

AU - Jones, Louis M.

AU - Neveu, Grégory

AU - Roussarie, Jean Pierre

AU - Rottier, Peter J M

AU - Tangy, Frédéric

AU - De Haan, Cornelis A M

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AB - Over the last 2 decades, yeast two-hybrid became an invaluable technique to decipher protein-protein interaction networks. In the field of virology, it has proven instrumental to identify virus-host interactions that are involved in viral embezzlement of cellular functions and inhibition of immune mechanisms. Here, we present a yeast two-hybrid protocol that has been used in our laboratory since 2006 to search for cellular partners of more than 300 viral proteins. Our aim was to develop a robust and straightforward pipeline, which minimizes false-positive interactions with a decent coverage of target cDNA libraries, and only requires a minimum of equipment. We also discuss reasons that motivated our technical choices and compromises that had to be made. This protocol has been used to screen most non-structural proteins of murine hepatitis virus (MHV), a member of betacoronavirus genus, against a mouse brain cDNA library. Typical results were obtained and are presented in this report.

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BT - Coronaviruses: Methods and Protocols

A2 - Maier, Helena Jane

A2 - Bickerton, Erica

A2 - Britton, Paul

PB - Springer New York

ER -

Vidalain PO, Jacob Y, Hagemeijer MC, Jones LM, Neveu G, Roussarie JP et al. A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions. In Maier HJ, Bickerton E, Britton P, editors, Coronaviruses: Methods and Protocols. Vol. V. Springer New York. 2015. p. 213-229. (Methods in Molecular Biology). doi: 10.1007/978-1-4939-2438-7_18

A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions

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