Diversity in genetic in vivo methods for protein-protein interaction studies

From the yeast two-hybrid system to the mammalian split-luciferase system

Bram Stynen, Helene Tournu, Jan Tavernier, Patrick Van Dijck

Research output: Contribution to journalReview article

100 Citations (Scopus)

Abstract

The yeast two-hybrid system pioneered the field of in vivo protein-rotein interaction methods and undisputedly gave rise to a palette of ingenious techniques that are constantly pushing further the limits of the original method. Sensitivity and selectivity have improved because of various technical tricks and experimental designs. Here we present an exhaustive overview of the genetic approaches available to study in vivo binary protein interactions, based on two-ybrid and protein fragment complementation assays. These methods have been engineered and employed successfully in microorganisms such as Saccharomyces cerevisiae and Escherichia coli, but also in higher eukaryotes. From single binary pairwise interactions to whole-genome interactome mapping, the self-reassembly concept has been employed widely. Innovative studies report the use of proteins such as ubiquitin, dihydrofolate reductase, and adenylate cyclase as reconstituted reporters. Protein fragment complementation assays have extended the possibilities in protein-protein interaction studies, with technologies that enable spatial and temporal analyses of protein complexes. In addition, one-hybrid and three-hybrid systems have broadened the types of interactions that can be studied and the findings that can be obtained. Applications of these technologies are discussed, together with the advantages and limitations of the available assays.

Original languageEnglish (US)
Pages (from-to)331-382
Number of pages52
JournalMicrobiology and Molecular Biology Reviews
Volume76
Issue number2
DOIs
StatePublished - Jun 1 2012

Fingerprint

Two-Hybrid System Techniques
Luciferases
Proteins
Spatio-Temporal Analysis
Technology
Tetrahydrofolate Dehydrogenase
Chromosome Mapping
Ubiquitin
Eukaryota
Adenylyl Cyclases
Saccharomyces cerevisiae
Research Design
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology
  • Immunology and Microbiology(all)
  • Infectious Diseases

Cite this

Diversity in genetic in vivo methods for protein-protein interaction studies : From the yeast two-hybrid system to the mammalian split-luciferase system. / Stynen, Bram; Tournu, Helene; Tavernier, Jan; Van Dijck, Patrick.

In: Microbiology and Molecular Biology Reviews, Vol. 76, No. 2, 01.06.2012, p. 331-382.

Research output: Contribution to journalReview article

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