Publication details

Optimized Detection of Protein-Protein and Protein-DNA Interactions, with Particular Application to Plant Telomeres

Authors

SCHOŘOVÁ Šárka FAJKUS Jiří PROCHÁZKOVÁ SCHRUMPFOVÁ Petra

Year of publication 2020
Type Chapter of a book
MU Faculty or unit

Faculty of Science

Citation
Description Characterization of protein-protein and protein-DNA interactions is critical to understand mechanisms governing the biology of cells. Here we describe optimized methods and their mutual combinations for this purpose: bimolecular fluorescence complementation (BiFC), co-immunoprecipitation (Co-IP), yeast two-hybrid systems (Y2H), and chromatin immunoprecipitation (ChIP). These improved protocols detect trimeric complexes in which two proteins of interest interact indirectly via a protein sandwiched between them. They also allow isolation of low-abundance chromatin proteins and confirmation that proteins of interest are associated with specific DNA sequences, for example telomeric tracts. Here we describe these methods and their application to map interactions of several telomere- and telomerase-associated proteins and to purify a sufficient amount of chromatin from Arabidopsis thaliana for further investigations (e.g., next-generation sequencing, hybridization)
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