Specificity determinants for the interaction of lambda repressor and P22 repressor dimers.
The related phage lambda and phage P22 repressors each bind cooperatively to adjacent and separated operator sites, an interaction that involves a pair of repressor dimers. The specificities of these interactions differ: Each dimer interacts with its own type but not with dimers of the heterologous repressor. The two repressors exhibit significant amino acid sequence homology in their carboxy-terminal domains, which are responsible for both dimer formation and the dimer-dimer interaction. Here, we identify a collection of amino acid substitutions that disrupt the protein-protein interaction of DNA-bound lambda repressor dimers and show that several of these substitutions have the same effect when introduced at the corresponding positions of P22 repressor. We use this information to construct a variant of the lambda repressor bearing only six non-wild-type amino acids that has a switched specificity; that is, it binds cooperatively with P22 repressor, but not with wild-type lambda repressor. These results identify a series of residues that determine the specificities of the two interactions.
Whipple
F W
FW
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115.
Kuldell
N H
NH
Cheatham
L A
LA
Hochschild
A
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D017124:Conserved Sequence; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009875:Operator Regions, Genetic / Q000235:genetics / Q000502:physiology; D017354:Point Mutation / Q000502:physiology; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology*; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins
D004279:DNA, Viral; D004268:DNA-Binding Proteins; C034261:P22 repressor protein, Enterobacteria phage P22; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins