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 (original) (raw)
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Gene, 1999
The nucleotide sequence of a DNA fragment previously shown to contain the attachment site (attP) of Oenococcus oeni phage fOg44 (Santos et al., 1998. Arch. Virol. 143, 523-536) has been determined. Sequence analysis indicated that this 6226 bp EcoRI fragment harbours an integrase gene, in the vicinity of a direct repeat rich region defining attP, as well as genes encoding a muramidase-related lysin (Lys) and a holin polypeptide (Hol). Transcriptional studies suggested that lys and hol are mainly co-expressed, late in the lytic cycle, from a promotor located upstream of lys. Between the lytic cassette and the phage integration elements three additional open reading frames were found: orf217 and orf252 of unknown function and orf72, the putative product of which bears 32% identity with acidic excisionases from other Gram positive phages. We have established that the first two orfs, as well as the predicted promotor of orf72, are included in a 2143-bp DNA segment missing from the genome of the deletion mutant fOg44D2. Although lysogens of fOg44 and fOg44D2 exhibited similar properties, each phage produced two distinguishable types of lysogenic strains, differing in inducibility and immunity to other oenophages.
Nisin-Triggered Activity of Lys44, the Secreted Endolysin from Oenococcus oeni Phage fOg44
Journal of Bacteriology, 2008
The intrinsic resistance of Oenococcus oeni cells to the secreted endolysin from oenophage fOg44 (Lys44) was investigated. Experiments with several antimicrobials support the hypothesis that the full activity of Lys44 requires sudden ion-nonspecific dissipation of the proton motive force, an event undertaken by the fOg44 holin in the phage infection context.
Enzymatic characterization of a lysin encoded by bacteriophage EL
Bacteriophage, 2013
The bacteriophage EL is a virus that specifically attacks the human pathogen Pseudomonas aeruginosa. This phage carries a large genome that encodes for its own chaperonin which presumably facilitates the proper folding of phage proteins independently of the host chaperonin system. EL also encodes a lysin enzyme, a critical component of the lytic cycle that is responsible for digesting the peptidoglycan layer of the host cell wall. Previously, this lysin was believed to be a substrate of the chaperonin encoded by phage EL. In order to characterize the activity of the EL lysin, and to determine whether lysin activity is contingent on chaperonin-mediated folding, a series of peptidoglycan hydrolysis activity assays were performed. Results indicate that the EL-encoded lysin has similar enzymatic activity to that of the Gallus gallus lysozyme and that the EL lysin folds into a functional enzyme in the absence of phage chaperonin and should not be considered a substrate.
FEMS Microbiology Letters, 1999
Malolactic fermentation by Oenococcus oeni is a crucial step in wine-making. Oe. oeni phages are thought to be responsible for fermentation failures, yet they have received little attention. After a molecular analysis concerning the phage P10MC integration system, this paper focuses on the lytic system. The attP (phage attachment site)-flanking region has been cloned and sequenced. The 1296-bp lysin gene (Lys) was identified in this region. The deduced amino acid sequence showed classical structural features of phage lysins, and this gene product expressed in Escherichia coli had a lytic activity against Oe. oeni. Downstream of Lys, a second ORF was present (P163). According to its amino acid sequence and the location of its gene, the product could be the P10MC holin. This study shows that the genomic organization of phage P10MC attP-flanking regions is very similar to that of other lactic acid bacteriophages.
Gene, 1992
Gene @A, which encodes lytic enzyme (LytA), of the isometric Lactococcus luctis bacteriophage $US3, was cloned and expressed in Escherichiu coli. The lytA gene was located on the physical map of the @US3 32-kb DNA that contains cohesive ends. Initial expression of &A was detected by lysis of an overlay of cells of the phage-sensitive strain, L. luctis SK1 12. However, LytA appeared to have a broad spectrum and induced lysis in more than 30 different lactococcal strains. The nucleotide sequence of &A showed a single open reading frame (ORF) of 774 bp encoding a protein of 258 amino acids (aa) with a calculated A4, of 28977. This is in agreement with the size of 29 kDa as determined for LytA produced in E. coli using a T7 expression system. The @A gene is preceded by an ORF that may code for a hydrophobic peptide of 66 aa containing a putative secretion signal, and two putative transmembrane helices. The deduced aa sequence of the phage @US3 LytA shows similarities to that of the autolysin of Streptococcus pneumoniue which is known to be an amidase.