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

Limin Shang1, Yuehui Zhang1, Yuchen Liu1

  • 1State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing 102206, China.

Genomics, Proteomics & Bioinformatics
|July 27, 2021
PubMed
Summary
This summary is machine-generated.

A new BiFC-seq method efficiently maps protein-protein interactions (PPIs) across the genome. This rapid technique identifies novel interactions for viruses and proteins like p53, advancing interactome mapping.

Keywords:
Bimolecular fluorescence complementationHigh-throughputNext-generation sequencingProtein–protein interaction

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biochemistry

Background:

  • Genome-wide protein-protein interaction (PPI) mapping is crucial but technologically challenging.
  • Existing methods often lack efficiency, scalability, or comprehensiveness.

Purpose of the Study:

  • To develop and validate a high-efficiency method for genome-wide interactome mapping.
  • To apply the new method to study viral and human protein interaction networks.

Main Methods:

  • Yeast-enhanced green fluorescent protein-based bimolecular fluorescence complementation (yEGFP-BiFC) was coupled with next-generation DNA sequencing (BiFC-seq).
  • The BiFC-seq method was applied to map the interactome of the Ebola virus and the tumor protein p53.
  • Pooled BiFC libraries were used to screen interactions in a complex biological background.

Main Results:

  • The BiFC-seq method successfully recaptured known Ebola virus (EBOV) interactions and identified five novel ones.
  • 97 interactors of p53 were identified, with over 75% being novel.
  • A network of 229 interactions among 205 proteins was revealed using pooled BiFC libraries.

Conclusions:

  • BiFC-seq is a highly sensitive, rapid, and economical method for comprehensive genome-wide interactome mapping.
  • This technology significantly advances the ability to discover and analyze protein interaction networks.