Induced Protein Dimerization in Vivo through Covalent Labeling

Abstract

We present here a novel approach to induce protein dimerization in living cells through covalent labeling of fusion proteins with ligands which are interacting with other proteins. The covalent labeling is based on the reaction of the DNA repair protein O⁶-alkylguanine-DNA alkyltransferase with O⁶-benzylguanine coupled to an appropriate ligand. Using methotrexate as a ligand, it is shown that the approach can be used to control transcription in the yeast Saccharomyces cerevisiae. The specificity of the labeling reaction and its irreversible nature should allow the approach to become a valuable tool to study and control protein dimerization in vivo.