PMID:22427494

Citation	Mayer, MJ, Gasson, MJ and Narbad, A (2012) Genomic sequence of bacteriophage ATCC 8074-B1 and activity of its endolysin and engineered variants against Clostridium sporogenes. Appl. Environ. Microbiol. 78:3685-92
Abstract	Lytic bacteriophage ATCC 8074-B1 produces large plaques on its host Clostridium sporogenes. Sequencing of the 47,595-bp genome allowed the identification of 82 putative open reading frames, including those encoding proteins for head and tail morphogenesis and lysis. However, sequences commonly associated with lysogeny were absent. ORF 22 encodes an endolysin, CS74L, that shows homology to N-acetylmuramoyl-L-alanine amidases, and when expressed in Escherichia coli, the protein causes effective lysis of C. sporogenes cells when added externally. CS74L was also active on Clostridium tyrobutyricum and Clostridium acetobutylicum. The catalytic domain expressed alone (CS74L(1-177)) exhibited a similar activity and the same host range as the full-length endolysin. A chimeric endolysin consisting of the CS74L catalytic domain fused to the C-terminal domain of endolysin CD27L, derived from Clostridium difficile bacteriophage ΦCD27, was produced. This chimera (CSCD) lysed C. sporogenes cells with an activity equivalent to that of the catalytic domain alone. In contrast, the CD27L C-terminal domain reduced the efficacy of the CS74L catalytic domain when tested against C. tyrobutyricum. The addition of the CD27L C-terminal domain did not enable the lysin to target C. difficile or other CD27L-sensitive bacteria.
Links	PubMed PMC3346354 Online version:10.1128/AEM.07884-11
Keywords	Bacteriophages/enzymology; Bacteriophages/genetics; Clostridium/virology; DNA, Viral/chemistry; DNA, Viral/genetics; Endopeptidases/genetics; Endopeptidases/metabolism; Genes, Viral; Genome, Viral; Molecular Sequence Data; Mutant Proteins/genetics; Mutant Proteins/metabolism; Open Reading Frames; Sequence Analysis, DNA; Spores, Bacterial/virology
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9CAUD:I1TJX3		GO:0044659: cytolysis by virus of host cell	ECO:0000314:		P		Figure 3 shows CS74L induced lysis of C. sporogenes 17786 cells in an E. coli expressing plasmid vector.	complete CACAO 10746
9CAUD:I1TJX3	involved_in	GO:0044659: cytolysis by virus of host cell	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete

References

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