Yeast protein-protein interaction assays and screens

Methods Mol Biol. 2011;754:145-65. doi: 10.1007/978-1-61779-154-3_8.

Abstract

Most transcription factors fulfill their role in protein complexes. As a consequence, information about their interaction capacity sheds light on a protein's function and the molecular mechanism underlying this activity. The yeast two-hybrid GAL4 (Y2H) assay is a powerful method to unravel and identify the composition of protein complexes. This in vivo based system makes use of two functional protein domains of the GAL4 transcription factor, each fused to a protein of interest. Upon interaction between the two proteins under study, a transcriptional activator gets reconstituted and reporter genes get activated, allowing the yeast to grow on selective medium. In this chapter protocols are given for Y2H library screening, directed Y2H screening, Y2H matrix screening, and YnH screening involving more than two proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Genes, Reporter
  • Multigene Family / genetics
  • Protein Binding / genetics
  • Protein Interaction Domains and Motifs
  • Protein Interaction Mapping / methods*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Two-Hybrid System Techniques*
  • Yeasts / genetics
  • Yeasts / growth & development
  • Yeasts / metabolism*

Substances

  • Fungal Proteins
  • Transcription Factors