Two-hybrid system

Definition

Two-hybrid systems are methods that assay for the interaction between two proteins, or protein and DNA in cells. Physical interaction is detected by the reconstitution of complementary domains of a reporter or transcription factor that activates reporter gene expression.

Latest Research and Reviews

  • Protocols |

    This protocol describes a nondestructive FRET-based assay for measuring protein interactions in cultured cells. The correlation between FRET efficiencies and relative protein concentration can be used to determine relative binding affinities.

    • Elisabeth S Butz
    • , Manu Ben-Johny
    • , Michael Shen
    • , Philemon S Yang
    • , Lingjie Sang
    • , Martin Biel
    • , David T Yue
    •  & Christian Wahl-Schott
    Nature Protocols 11, 2470–2498
  • Research | | open

    The centrosome is a large intracellular structure that serves as the microtubule-organising center, but how it is accurately assembled is not known. Here the authors generate a ‘domain-level’ centrosome interactome and show that Plk4 positions the essential centriole component Asterless by phosphorylating Cep135.

    • Brian J. Galletta
    • , Carey J. Fagerstrom
    • , Todd A. Schoborg
    • , Tiffany A. McLamarrah
    • , John M. Ryniawec
    • , Daniel W. Buster
    • , Kevin C. Slep
    • , Gregory C. Rogers
    •  & Nasser M. Rusan
  • Research |

    A high-sensitivity, high-quality yeast two-hybrid screen using short-read sequencing as its readout is used to map the interactome of human methyltransferases.

    • Mareike Weimann
    • , Arndt Grossmann
    • , Jonathan Woodsmith
    • , Ziya Özkan
    • , Petra Birth
    • , David Meierhofer
    • , Nouhad Benlasfer
    • , Taras Valovka
    • , Bernd Timmermann
    • , Erich E Wanker
    • , Sascha Sauer
    •  & Ulrich Stelzl
    Nature Methods 10, 339–342
  • Research |

    The authors describe the enhanced yeast one-hybrid platform for large-scale screening of protein-DNA interactions and test its performance by mapping Caenorhabditis elegans gene regulatory networks. Also in this issue, Hens et al. describe an alternative platform for this purpose and apply it to screen for transcription factor–DNA interactions in Drosophila melanogaster.

    • John S Reece-Hoyes
    • , Alos Diallo
    • , Bryan Lajoie
    • , Amanda Kent
    • , Shaleen Shrestha
    • , Sreenath Kadreppa
    • , Colin Pesyna
    • , Job Dekker
    • , Chad L Myers
    •  & Albertha J M Walhout
    Nature Methods 8, 1059–1064
  • Research |

    An almost-complete, sequence-verified collection of Arabidopsis thaliana root stele transcription factors is reported. The authors use it in the enhanced yeast-one hybrid (eY1H) assay to map gene regulatory interactions in the plant. Also in this issue, Reece-Hoyes et al. describe the eY1H pipeline.

    • Allison Gaudinier
    • , Lifang Zhang
    • , John S Reece-Hoyes
    • , Mallorie Taylor-Teeples
    • , Li Pu
    • , Zhijie Liu
    • , Ghislain Breton
    • , Jose L Pruneda-Paz
    • , Dahae Kim
    • , Steve A Kay
    • , Albertha J M Walhout
    • , Doreen Ware
    •  & Siobhan M Brady
    Nature Methods 8, 1053–1055

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