Induced protein dimerization in vivo through covalent labeling

J Am Chem Soc. 2003 Dec 10;125(49):14970-1. doi: 10.1021/ja037883p.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Dimerization
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Glutathione Transferase / biosynthesis
  • Glutathione Transferase / chemistry
  • Glutathione Transferase / genetics
  • Guanine / analogs & derivatives*
  • Guanine / chemistry
  • Guanine / metabolism*
  • Humans
  • Methotrexate / chemistry
  • Methotrexate / metabolism*
  • O(6)-Methylguanine-DNA Methyltransferase / biosynthesis*
  • O(6)-Methylguanine-DNA Methyltransferase / chemistry
  • O(6)-Methylguanine-DNA Methyltransferase / genetics
  • Receptors, Glucocorticoid / biosynthesis
  • Receptors, Glucocorticoid / chemistry
  • Receptors, Glucocorticoid / genetics
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Serine Endopeptidases / biosynthesis
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / genetics
  • Tetrahydrofolate Dehydrogenase / biosynthesis
  • Tetrahydrofolate Dehydrogenase / chemistry
  • Tetrahydrofolate Dehydrogenase / genetics
  • Transcriptional Activation / drug effects
  • Yeasts / enzymology
  • Yeasts / genetics
  • Yeasts / metabolism

Substances

  • Bacterial Proteins
  • LexA protein, Bacteria
  • Receptors, Glucocorticoid
  • Recombinant Fusion Proteins
  • O(6)-benzylguanine
  • Guanine
  • Tetrahydrofolate Dehydrogenase
  • O(6)-Methylguanine-DNA Methyltransferase
  • Glutathione Transferase
  • Serine Endopeptidases
  • Methotrexate