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

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
Subtitle of host publication	Methods and Protocols
Editors	Helena Jane Maier, Erica Bickerton, Paul Britton
Publisher	Springer
Chapter	18
Pages	213-229
Number of pages	17
ISBN (Electronic)	978-1-4939-2438-7
ISBN (Print)	978-1-4939-2437-0
DOIs	https://doi.org/10.1007/978-1-4939-2438-7_18
Publication status	Published - 2015

Publication series

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

Keywords

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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

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

Vidalain2015_Protocol_AField-ProvenYeastTwo-HybridPrFinal published version, 832 KBLicence: Taverne

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 (pp. 213-229). (Methods in Molecular Biology; Vol. 1282). Springer. https://doi.org/10.1007/978-1-4939-2438-7_18

@inbook{47950da77fd2459da84025e9b772228c,

title = "A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions",

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

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

year = "2015",

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

language = "English",

isbn = "978-1-4939-2437-0",

series = "Methods in Molecular Biology",

publisher = "Springer",

pages = "213--229",

editor = "Maier, {Helena Jane } and Bickerton, {Erica } and Britton, {Paul }",

booktitle = "Coronaviruses",

}

Vidalain, P-O, Jacob, Y, Hagemeijer, MC, Jones, LM, Neveu, G, Roussarie, J-P, 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. Methods in Molecular Biology, vol. 1282, Springer, 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. Springer, 2015. p. 213-229 (Methods in Molecular Biology; Vol. 1282).

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

TY - CHAP

T1 - A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions

AU - Vidalain, Pierre-Olivier

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

PY - 2015

Y1 - 2015

N2 - 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.

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.

KW - Murine hepatitis virus

KW - Host–pathogen interactions

KW - Yeast two-hybrid

KW - Interactomics

KW - Proteomics

U2 - 10.1007/978-1-4939-2438-7_18

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

M3 - Chapter

C2 - 25720483

SN - 978-1-4939-2437-0

T3 - Methods in Molecular Biology

SP - 213

EP - 229

BT - Coronaviruses

A2 - Maier, Helena Jane

A2 - Bickerton, Erica

A2 - Britton, Paul

PB - Springer

ER -

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

Abstract

Publication series

Keywords

UN SDGs

Access to Document

Fingerprint

Cite this