The N-terminal region of the Oenococcus oeni bacteriophage fOg44 lysin behaves as a bona fide signal peptide in Escherichia coli and as a cis-inhibitory element, preventing lytic activity on oenococcal cells.
The function of the N-terminal region of the Oenococcus oeni phage fOg44 lysin (Lys44) as an export signal was investigated. We observed that when induced in Escherichia coli, Lys44 was cleaved between residues 27 and 28 in a SecA-dependent manner. Lys44 processing could be blocked by a specific signal peptidase inhibitor and was severely reduced by modification of the cleavage site. The lethal effect of Lys44 expression observed in E. coli was ascribed to the presence of its N-terminal 27-residue sequence, as its deletion resulted in the production of a nontoxic, albeit active, product. We have further established that lytic activity in oenococcal cells was dependent on Lys44 processing. An active protein with the molecular mass expected for the cleaved enzyme was detected in extracts from O. oeni-infected cells. The temporal pattern of its appearance suggests that synthesis and export of Lys44 in the infected host progress along with phage maturation. Overall, these results provide, for the first time, experimental evidence for the presence of a signal peptide in a bacteriophage lysin. Database searches and alignment of protein sequences support the prediction that other known O. oeni and Lactococcus lactis phages also encode secretory lysins. The evolutionary significance of a putative phage lysis mechanism relying on secretory lytic enzymes is tentatively discussed, on the basis of host cell wall structure and autolytic capacity.
São-José
C
C
Centro de Genética e Biologia Molecular e Departamento de Biologia Vegetal, Faculdade de Ciências da Universidade de Lisboa, 1700 Lisbon, Portugal.
Parreira
R
R
Vieira
G
G
Santos
M A
MA
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000378:metabolism*; D001483:Base Sequence; D002473:Cell Wall / Q000201:enzymology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism* / Q000821:virology; D006095:Gram-Positive Cocci / Q000821:virology*; D006867:Hydrolases / Q000378:metabolism; D007700:Kinetics; D007934:Leuconostoc / Q000821:virology; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
C117327:Lys protein, Oenococcus oeni temperate bacteriophage fOg44; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D014764:Viral Proteins; D006867:Hydrolases