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Nature Biotechnology 22, 888–892 (1 July 2004) | doi:10.1038/nbt985

A tethered catalysis, two-hybrid system to identify protein-protein interactions requiring post-translational modifications

Dawei Guo , Tony R Hazbun , Xin-Jing Xu , Sze-Ling Ng , Stanley Fields & Min-Hao Kuo

We have modified the yeast two-hybrid system to enable the detection of protein-protein interactions that require a specific post-translational modification, using the acetylation of histones and the phosphorylation of the carboxyl terminal domain (CTD) of RNA polymerase II as test modifications. In this tethered catalysis assay, constitutive modification of the protein to be screened for interactions is achieved by fusing it to its cognate modifying enzyme, with the physical linkage resulting in efficient catalysis.