title: The lytic enzyme of the pneumococcal phage Dp-1: a chimeric lysin of intergeneric origin.
abstract: We have localized, cloned and characterized the genes coding for the lytic system of the pneumococcal phage Dp-1. The lytic enzyme of this phage (Pal), previously identified as an N-acetyl-muramoyl-L-alanine amidase, shows a modular organization similar to that described for the lytic enzymes of Streptococcus pneumoniae and its bacteriophages. The construction of chimeric enzymes between pneumococcus and bacteria (or phages) that belong to different Gram-positive families has shown that the interchange of functional domains switches enzyme specificity. Interestingly, Pal appears to be a natural chimeric enzyme of intergeneric origin, that is the N-terminal domain was highly similar to that of the murein hydrolase coded by a gene found in the phage BK5-T that infects Lactococcus lactis, whereas the C-terminal domain was homologous to those found in the lytic enzymes of the pneumococcal system that is responsible for the binding to the choline residues present in the cell wall substrate. Biochemical analysis of Pal revealed that this enzyme shares important properties with those of the major LytA101 autolysin found in an atypical, clinical pneumococcal isolate. These peculiar characteristics have been ascribed to a modified C-terminal domain. The natural chimeric enzyme described here provides further support for the theory of modular evolution of proteins and its characteristics also furnish interesting clues on the molecular mechanisms involved in the more invasive types of atypical pneumococci.
authors.affiliation: Departamento de Microbiología Molecular, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Cientificas, Velázquez, Madrid, Spain.
authors.forename: M M
authors.initials: MM
authors.lastname: Sheehan
authors.forename: J L
authors.initials: JL
authors.lastname: García
authors.forename: R
authors.initials: R
authors.lastname: López
authors.forename: P
authors.initials: P
authors.lastname: García
mesh_terms: D000595:Amino Acid Sequence; D002794:Choline / Q000378:metabolism / Q000494:pharmacology; D003001:Cloning, Molecular; D015342:DNA Probes; D004591:Electrophoresis, Polyacrylamide Gel; D013294:Lactococcus lactis / Q000821:virology; D008242:Lysogeny / Q000235:genetics; D008969:Molecular Sequence Data; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016366:Open Reading Frames; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins; D015183:Restriction Mapping; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D017108:Streptococcus Phages / Q000201:enzymology* / Q000235:genetics; D013296:Streptococcus pneumoniae / Q000821:virology
chemical_list: D015342:DNA Probes; D011993:Recombinant Fusion Proteins; C076332:EJL amidase; D009238:N-Acetylmuramoyl-L-alanine Amidase; D002794:Choline