title: Reassortment of DNA recognition domains and the evolution of new specificities.
abstract: Type I restriction enzymes comprise three subunits only one of which, the S polypeptide, dictates the specificity of the DNA sequence recognized. Recombination between two different hsdS genes, SP and SB, led to the isolation of a system, SQ, which had a different specificity from that of either parent. The finding that the nucleotide sequence recognized by SQ is a hybrid containing components from both the SP and SB target sequences suggested that DNA recognition is carried out by two separable domains within each specificity polypeptide. To test this we have made the recombinant gene of reciprocal structure and demonstrate that it encodes a polypeptide whose recognition sequence, deduced in vivo, is as predicted by this model. We also report the sequence of the SB specificity gene, so that information is now available for the five known members of this family of enzymes. All show a similar organization of conserved and variable regions. Comparisons of the predicted amino acid sequences reveal large non-conserved areas which may not even be structurally similar. This is remarkable since these different S subunits are functionally identical, except for the specificity with respect to the DNA sequence with which they interact. We discuss the correlation of the variation in polypeptide sequence with recognition specificities.
authors.affiliation: Department of Molecular Biology, University of Edinburgh, UK.
authors.forename: A A
authors.initials: AA
authors.lastname: Gann
authors.forename: A J
authors.initials: AJ
authors.lastname: Campbell
authors.forename: J F
authors.initials: JF
authors.lastname: Collins
authors.forename: A F
authors.initials: AF
authors.lastname: Coulson
authors.forename: N E
authors.initials: NE
authors.lastname: Murray
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics*; D001483:Base Sequence; D005075:Biological Evolution; D004262:DNA Restriction Enzymes / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D005814:Genes, Viral; D046911:Macromolecular Substances; D008969:Molecular Sequence Data
chemical_list: D046911:Macromolecular Substances; D004262:DNA Restriction Enzymes