title: Next-Generation Sequencing for Binary Protein-Protein Interactions.
abstract: The yeast two-hybrid (Y2H) system exploits host cell genetics in order to display binary protein-protein interactions (PPIs) via defined and selectable phenotypes. Numerous improvements have been made to this method, adapting the screening principle for diverse applications, including drug discovery and the scale-up for proteome wide interaction screens in human and other organisms. Here we discuss a systematic workflow and analysis scheme for screening data generated by Y2H and related assays that includes high-throughput selection procedures, readout of comprehensive results via next-generation sequencing (NGS), and the interpretation of interaction data via quantitative statistics. The novel assays and tools will serve the broader scientific community to harness the power of NGS technology to address PPI networks in health and disease. We discuss examples of how this next-generation platform can be applied to address specific questions in diverse fields of biology and medicine.
authors.affiliation: Next Interactions, Inc., Richmond CA, USA.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Suter
authors.affiliation: BioInfoRx, Inc., Madison WI, USA.
authors.forename: Xinmin
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Next Interactions, Inc., Richmond CA, USA.
authors.forename: C Gustavo
authors.initials: CG
authors.lastname: Pesce
authors.affiliation: Next Interactions, Inc., RichmondCA, USA; Regenerative Sciences Institute, SunnyvaleCA, USA.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Mendelsohn
authors.affiliation: Department of Plant Biology, University of California, Davis, Davis CA, USA.
authors.forename: Savithramma P
authors.initials: SP
authors.lastname: Dinesh-Kumar
authors.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley CA, USA.
authors.forename: Jian-Hua
authors.initials: JH
authors.lastname: Mao
keywords: interactome mapping; next-generation sequencing; protein–protein interactions; quantitative interaction profiles; yeast two-hybrid