The power of the yeast two-hybrid system in the identification of novel drug targets: building and modulating PPP1 interactomes

Abstract

Since the description of the yeast two-hybrid (Y2H) method, it has become more and more evident that it is the most commonly used method to identify protein–protein interactions (PPIs). The improvements in the original Y2H methodology in parallel with the idea that PPIs are promising drug targets, offer an excellent opportunity to apply the principles of this molecular biology technique to the pharmaceutical field. Additionally, the theoretical developments in the networks field make PPI networks very useful frameworks that facilitate many discoveries in biomedicine. This review highlights the relevance of Y2H in the determination of PPIs, specifically phosphoprotein phosphatase 1 interactions, and its possible outcomes in pharmaceutical research.

Keywords:

• drug screening
• drug targets
• interactomes
• molecular interactions
• phosphoprotein phosphatase 1
• protein–protein interactions
• yeast two-hybrid variants

Financial & competing interests disclosure

This work was supported by the Fundação para a Ciência e Tecnologia (FCT; PTDC/DTP-PIC/0460/2012) and cofinanced by FEDER through ‘Eixo I do Programa Operacional Fatores de Competitividade (POFC) do QREN’ (COMPETE: FCOMP-01–0124-FEDER-028692). JV Silva is a recipient of a PhD fellowship from FCT (SFRH/BD/81458/2011); MJ Freitas is a recipient of a PhD fellowship from FCT (SFRH/BD/84876/2012) and J Felgueiras is a recipient of research fellowship from the FCT project PTDC/DTP-PIC/0460/2012, cofinanced by FEDER through ‘Eixo I do Programa Operacional Fatores de Competitividade (POFC) do QREN’ (COMPETE: FCOMP-01–0124-FEDER-028692). The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending or royalties.

Key issues

• The disadvantages of the conventional yeast two-hybrid (Y2H) system led to several adaptations, which gave rise to variations of the original technique.

• The high-throughput Y2H screening was the most significant improvement of this technique and has been essential to unravel molecular interactions, namely protein–protein interactions.

• The Y2H technique proved to be particularly critical in the comprehension of phosphoprotein phosphatase 1 (PPP1) molecular roles.

• PPP1 function depends on its multiple targeting/regulatory subunits, known as PPP1-interacting proteins (PIPs), many of which were identified by Y2H.

• The application of the Y2H system in drug discovery is a recent but very promising phenomenon. Several variants of the method have been particularly adapted aiming pharmacological studies, including the reverse Y2H screening and the yeast three-hybrid.

• PPP1 complexes represent promising therapeutic targets and the Y2H represents a useful tool to identify PPP1 complexes-modulating drugs.

Notes