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Reference: Moreno D, et al. (2013) A fluorescent reporter for mapping cellular protein-protein interactions in time and space. Mol Syst Biol 9:647

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Abstract

We introduce a fluorescent reporter for monitoring protein-protein interactions in living cells. The method is based on the Split-Ubiquitin method and uses the ratio of two auto-fluorescent reporter proteins as signal for interaction (SPLIFF). The mating of two haploid yeast cells initiates the analysis and the interactions are followed online by two-channel time-lapse microscopy of the diploid cells during their first cell cycle. Using this approach we could with high spatio-temporal resolution visualize the differences between the interactions of the microtubule binding protein Stu2p with two of its binding partners, monitor the transient association of a Ran-GTPase with its receptors at the nuclear pore, and distinguish between protein interactions at the polar cortical domain at different phases of polar growth. These examples further demonstrate that protein-protein interactions identified from large-scale screens can be effectively followed up by high-resolution single-cell analysis.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't
Authors
Moreno D, Neller J, Kestler HA, Kraus J, Dunkler A, Johnsson N
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