PMID:26683375

Citation	Dunne, M, Leicht, S, Krichel, B, Mertens, HD, Thompson, A, Krijgsveld, J, Svergun, DI, Gómez-Torres, N, Garde, S, Uetrecht, C, Narbad, A, Mayer, MJ and Meijers, R (2016) Crystal Structure of the CTP1L Endolysin Reveals How Its Activity Is Regulated by a Secondary Translation Product. J. Biol. Chem. 291:4882-93
Abstract	Bacteriophages produce endolysins, which lyse the bacterial host cell to release newly produced virions. The timing of lysis is regulated and is thought to involve the activation of a molecular switch. We present a crystal structure of the activated endolysin CTP1L that targets Clostridium tyrobutyricum, consisting of a complex between the full-length protein and an N-terminally truncated C-terminal cell wall binding domain (CBD). The truncated CBD is produced through an internal translation start site within the endolysin gene. Mutants affecting the internal translation site change the oligomeric state of the endolysin and reduce lytic activity. The activity can be modulated by reconstitution of the full-length endolysin-CBD complex with free CBD. The same oligomerization mechanism applies to the CD27L endolysin that targets Clostridium difficile and the CS74L endolysin that targets Clostridium sporogenes. When the CTP1L endolysin gene is introduced into the commensal bacterium Lactococcus lactis, the truncated CBD is also produced, showing that the alternative start codon can be used in other bacterial species. The identification of a translational switch affecting oligomerization presented here has implications for the design of effective endolysins for the treatment of bacterial infections.
Links	PubMed PMC4777826 Online version:10.1074/jbc.M115.671172
Keywords	Amino Acid Sequence; Bacteriophages/enzymology; Bacteriophages/genetics; Clostridium tyrobutyricum/drug effects; Codon, Initiator; Endopeptidases/chemistry; Endopeptidases/genetics; Endopeptidases/metabolism; Endopeptidases/toxicity; Molecular Sequence Data; Mutation; Protein Binding; Protein Multimerization
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9CAUD:D9ZNF3		GO:0009253: peptidoglycan catabolic process	ECO:0000315:		P		In figure 4 they show a sequence alignment on the nucleotide level of CTP1L and CS74L. In the figure there are other related endolysins shown that are present. The putative Shine-Dalgarno region is shown in figure 4a and 4b in the red. It also indicates that the start codon is GTG. The nucleotide sequence used during E. coli expression of when they did the sequence alignment was derived from the original bacteriophage DNA. To test whether the wild-type sequence encoded a secondary translation site to the wild type.	complete CACAO 12880

References

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