Control of gene expression with small molecules: biotin-mediated acylation of targeted lysine residues in recombinant yeast.
Chemical inducers of dimerization (CIDs) are powerful tools for controlling diverse cellular processes. These small molecules typically form strong noncovalent interactions with proteins. We report a related approach involving covalent acylation of a specific lysine residue of a target protein by the small molecule biotin. To control protein-protein interactions with biotin, the biotin protein ligase BirA from E. coli was coexpressed in yeast with a streptavidin-LexA fusion protein and Avitag or BCCP biotin acceptor peptides fused to the B42 activation domain. The addition of biotin (10 nM) resulted in BirA-mediated biotinylation of the biotin acceptor protein, recruitment to LexA DNA sites, and maximal activation of reporter gene expression in this yeast tribrid system. The high potency, low toxicity, and low molecular weight of biotin as a covalent CID are attractive properties for controlling cellular processes.
Athavankar
Sonalee
S
Department of Chemistry, The Pennsylvania State University, 152 Davey Laboratory, University Park, PA 16802, USA.
Peterson
Blake R
BR
D000103:Acetyl-CoA Carboxylase / Q000235:genetics / Q000378:metabolism; D000215:Acylation / Q000187:drug effects; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001710:Biotin / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D019731:Carbon-Nitrogen Ligases / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D054889:Fatty Acid Synthase, Type II; D005786:Gene Expression Regulation / Q000187:drug effects*; D008239:Lysine / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000099160:LexA Repressor Protein
D000103:Acetyl-CoA Carboxylase; D001426:Bacterial Proteins; D001710:Biotin; D019731:Carbon-Nitrogen Ligases; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D054889:Fatty Acid Synthase, Type II; D008239:Lysine; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D012697:Serine Endopeptidases; D014157:Transcription Factors; D000099160:LexA Repressor Protein; C058125:birA protein, E coli; C079633:biotin carboxyl carrier protein