Induced protein dimerization in vivo through covalent labeling.
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.
Gendreizig
Susanne
S
Institute of Molecular and Biological Chemistry, Institute of Biomolecular Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland.
Kindermann
Maik
M
Johnsson
Kai
K
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D006147:Guanine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008727:Methotrexate / Q000737:chemistry / Q000378:metabolism*; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011965:Receptors, Glucocorticoid / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects; D015003:Yeasts / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D000099160:LexA Repressor Protein
D001426:Bacterial Proteins; D005982:Glutathione Transferase; D006147:Guanine; D008727:Methotrexate; D019853:O(6)-Methylguanine-DNA Methyltransferase; D011965:Receptors, Glucocorticoid; D011993:Recombinant Fusion Proteins; D012697:Serine Endopeptidases; D013762:Tetrahydrofolate Dehydrogenase; D000099160:LexA Repressor Protein; C064976:O(6)-benzylguanine