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A Yeast BiFC-seq Method for Genome-wide Interactome Mapping

Genome-wide physical protein–protein interaction (PPI) mapping remains a major challenge for current technologies. Here, we reported a high-efficiency BiFC-seq method, yeast-enhanced green fluorescent protein-based bimolecular fluorescence complementation (yEGFP-BiFC) coupled with next-generation DN...

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Autores principales: Shang, Limin, Zhang, Yuehui, Liu, Yuchen, Jin, Chaozhi, Yuan, Yanzhi, Tian, Chunyan, Ni, Ming, Bo, Xiaochen, Zhang, Li, Li, Dong, He, Fuchu, Wang, Jian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9880813/
https://www.ncbi.nlm.nih.gov/pubmed/34314873
http://dx.doi.org/10.1016/j.gpb.2021.02.008
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author Shang, Limin
Zhang, Yuehui
Liu, Yuchen
Jin, Chaozhi
Yuan, Yanzhi
Tian, Chunyan
Ni, Ming
Bo, Xiaochen
Zhang, Li
Li, Dong
He, Fuchu
Wang, Jian
author_facet Shang, Limin
Zhang, Yuehui
Liu, Yuchen
Jin, Chaozhi
Yuan, Yanzhi
Tian, Chunyan
Ni, Ming
Bo, Xiaochen
Zhang, Li
Li, Dong
He, Fuchu
Wang, Jian
author_sort Shang, Limin
collection PubMed
description Genome-wide physical protein–protein interaction (PPI) mapping remains a major challenge for current technologies. Here, we reported a high-efficiency BiFC-seq method, yeast-enhanced green fluorescent protein-based bimolecular fluorescence complementation (yEGFP-BiFC) coupled with next-generation DNA sequencing, for interactome mapping. We first applied yEGFP-BiFC method to systematically investigate an intraviral network of the Ebola virus. Two-thirds (9/14) of known interactions of EBOV were recaptured, and five novel interactions were discovered. Next, we used the BiFC-seq method to map the interactome of the tumor protein p53. We identified 97 interactors of p53, more than three-quarters of which were novel. Furthermore, in a more complex background, we screened potential interactors by pooling two BiFC libraries together and revealed a network of 229 interactions among 205 proteins. These results show that BiFC-seq is a highly sensitive, rapid, and economical method for genome-wide interactome mapping.
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spelling pubmed-98808132023-01-28 A Yeast BiFC-seq Method for Genome-wide Interactome Mapping Shang, Limin Zhang, Yuehui Liu, Yuchen Jin, Chaozhi Yuan, Yanzhi Tian, Chunyan Ni, Ming Bo, Xiaochen Zhang, Li Li, Dong He, Fuchu Wang, Jian Genomics Proteomics Bioinformatics Method Genome-wide physical protein–protein interaction (PPI) mapping remains a major challenge for current technologies. Here, we reported a high-efficiency BiFC-seq method, yeast-enhanced green fluorescent protein-based bimolecular fluorescence complementation (yEGFP-BiFC) coupled with next-generation DNA sequencing, for interactome mapping. We first applied yEGFP-BiFC method to systematically investigate an intraviral network of the Ebola virus. Two-thirds (9/14) of known interactions of EBOV were recaptured, and five novel interactions were discovered. Next, we used the BiFC-seq method to map the interactome of the tumor protein p53. We identified 97 interactors of p53, more than three-quarters of which were novel. Furthermore, in a more complex background, we screened potential interactors by pooling two BiFC libraries together and revealed a network of 229 interactions among 205 proteins. These results show that BiFC-seq is a highly sensitive, rapid, and economical method for genome-wide interactome mapping. Elsevier 2022-08 2021-07-24 /pmc/articles/PMC9880813/ /pubmed/34314873 http://dx.doi.org/10.1016/j.gpb.2021.02.008 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Method
Shang, Limin
Zhang, Yuehui
Liu, Yuchen
Jin, Chaozhi
Yuan, Yanzhi
Tian, Chunyan
Ni, Ming
Bo, Xiaochen
Zhang, Li
Li, Dong
He, Fuchu
Wang, Jian
A Yeast BiFC-seq Method for Genome-wide Interactome Mapping
title A Yeast BiFC-seq Method for Genome-wide Interactome Mapping
title_full A Yeast BiFC-seq Method for Genome-wide Interactome Mapping
title_fullStr A Yeast BiFC-seq Method for Genome-wide Interactome Mapping
title_full_unstemmed A Yeast BiFC-seq Method for Genome-wide Interactome Mapping
title_short A Yeast BiFC-seq Method for Genome-wide Interactome Mapping
title_sort yeast bifc-seq method for genome-wide interactome mapping
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9880813/
https://www.ncbi.nlm.nih.gov/pubmed/34314873
http://dx.doi.org/10.1016/j.gpb.2021.02.008
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