Two-hybrid system

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 |

    A sequence-verified collection of human transcription factors is reported. The authors used it in the enhanced yeast-one hybrid (eY1H) assay to map human gene regulatory networks. Also in this issue, Reece-Hoyes et al. describe the eY1H pipeline.

    • John S Reece-Hoyes
    • , A Rasim Barutcu
    • , Rachel Patton McCord
    • , Jun Seop Jeong
    • , Lizhi Jiang
    • , Andrew MacWilliams
    • , Xinping Yang
    • , Kourosh Salehi-Ashtiani
    • , David E Hill
    • , Seth Blackshaw
    • , Heng Zhu
    • , Job Dekker
    •  & Albertha J M Walhout
    Nature Methods 8, 1050-1052
  • 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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