Cloning and expression in Escherichia coli and Staphylococcus aureus of the beta-lysin determinant from Staphylococcus aureus: evidence that bacteriophage conversion of beta-lysin activity is caused by insertional inactivation of the beta-lysin determinant

Coleman, David C.; Arbuthnott, J. P.; Pomeroy, H.; Birkbeck, T. H.

doi:10.1016/0882-4010(86)90040-9

Cited by 76 publications

(48 citation statements)

References 42 publications

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“…None of the 17 communityacquired-septicaemia strains produced P-lysin. This may be due to a higher incidence of carriage of P-lysin/staphylokinase double-converting prophages (Coleman et al, 1986). In support of this hypothesis, the frequency of staphylokinase production by hospital-acquired isolates (74%) was significantly (p < 0.01) lower than that of the community-acquired strains (94%).…”

Section: Discussionmentioning

confidence: 73%

Enterotoxin production by Staphylococcus aureus isolates from cases of septicaemia and from healthy carriers

Humphreys

Keane

Hone

et al. 1989

Journal of Medical Microbiology

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Section: Discussionmentioning

confidence: 73%

Enterotoxin production by Staphylococcus aureus isolates from cases of septicaemia and from healthy carriers

Humphreys

Keane

Hone

et al. 1989

Journal of Medical Microbiology

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“…Over the past decade, several microbial SMases have been cloned and characterized (4,7,21,30). The amino acid sequence of TK4 SMase shows about 30 to 40% identity with that of other microbial SMases reported to date; however, little identity was found with mammalian SMases.…”

Section: Discussionmentioning

confidence: 99%

Molecular Cloning and Expression of Mn ²⁺ -Dependent Sphingomyelinase/Hemolysin of an Aquatic Bacterium, Pseudomonas sp. Strain TK4

et al. 2002

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We report here the molecular cloning and expression of a hemolytic sphingomyelinase from an aquatic bacterium, Pseudomonas sp. strain TK4. The sphingomyelinase gene was found to consist of 1,548 nucleotides encoding 516 amino acid residues. The recombinant 57.7-kDa enzyme hydrolyzed sphingomyelin but not phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, or phosphatidylethanolamine, indicating that the enzyme is a sphingomyelin-specific sphingomyelinase C. The hydrolysis of sphingomyelin by the enzyme was found to be most efficient at pH 8.0 and activated by Mn 2؉ . The enzyme shows quite a broad specificity, i.e., it hydrolyzed 4-nitrobenz-2-oxa-1,3-diazole (NBD)-sphingomyelin with shortchain fatty acids and NBD-sphingosylphosphorylcholine, the latter being completely resistant to hydrolysis by any sphingomyelinase reported so far. Significant sequence similarities were found in sphingomyelinases from Bacillus cereus, Staphylococcus aureus, Listeria ivanovii, and Leptospira interrogans, as well as a hypothetical protein encoded in Chromobacterium violaceum, although the first three lacked one-third of the sequence corresponding to that from the C terminus of the TK4 enzyme. Interestingly, the deletion mutant of strain TK4 lacking 186 amino acids at the C-terminal end hydrolyzed sphingomyelin, whereas it lost all hemolytic activity, indicating that the C-terminal region of the TK4 enzyme is indispensable for the hemolytic activity.

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“…RN1 carries three prophages (⌽11, ⌽12, and ⌽13) (21,30). It is agr group 1 and agr ϩ , and it produces alpha-, delta-, and gamma-hemolysins, but no beta-hemolysin, as prophage ⌽13 is integrated in hlb (10). The 8325 lineage remains the standard for basic research on gene regulation, plasmid replication, prophage behavior, cell wall structures, membrane composition, physiology, and antibiotic resistance.…”

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confidence: 99%

Repair of Global Regulators inStaphylococcus aureus8325 and Comparative Analysis with Other Clinical Isolates

et al. 2010

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The pathogenicity of Staphylococcus aureus strains varies tremendously (as seen with animals). It is largely dependent on global regulators, which control the production of toxins, virulence, and fitness factors. Despite the vast knowledge of staphylococcal molecular genetics, there is still widespread dispute over what factors must come together to make a strain highly virulent. S. aureus NCTC8325 (RN1 and derivatives) is a widely used model strain for which an incomparable wealth of knowledge has accumulated in the almost 50 years since its isolation. Although RN1 has functional agr, sarA, and sae global regulators, it is defective in two regulatory genes, rsbU (a positive activator of SigB) and tcaR (an activator of protein A transcription), and is therefore considered by many to be a poor model for studies of regulation and virulence. Here, we repaired these genes and compared the resulting RN1 derivatives with other widely used strains, Newman, USA300, UAMS-1, and COL, plus the parental RN1, with respect to growth, extracellular protein pattern, hemolytic activity, protein A production, pigmentation, biofilm formation, and mouse lethality. The tcaR-repaired strain, showed little alteration in these properties. However, the rsbU-repaired strain was profoundly altered. Hemolytic activity was largely decreased, the exoprotein pattern became much more similar to that of typical wild-type (wt) S. aureus, and there was a surprising increase in mouse lethality. We note that each of the strains tested has a mutational alteration in one or more other regulatory functions, and we conclude that the repaired RN1 is a good model strain for studies of staphylococcal regulation and pathobiology; although strain Newman has been used extensively for such studies in recent years, it has a missense mutation in saeS, the histidine kinase component of the sae signaling module, which profoundly alters its regulatory phenotype. If this mutation were repaired, Newman would be considerably improved as a model strain.As a classical, dangerous, and universal human pathogen, Staphylococcus aureus has aroused continuing interest in its epidemiology, pathogenesis, and antibiotic resistance and other features of its pathobiology. This interest has led, concomitantly with the development of bacterial molecular biology, to the development of a model strain for the analysis of staphylococcal molecular genetics in relation to the pathogenicity of the organism. This strain, NCTC8325, was isolated in 1960 from a sepsis patient and utilized as the propagating strain for phage 47 of the international phage typing system. It was originally chosen for research owing to its sensitivity to all known antibiotics and was initially used primarily for studies of antibiotic resistance transfer and carriage by plasmids (32). It is designated RN1 in the Novick lab strain collection and is presently maintained by the Central Public Health Laboratory, Colindale, London, United Kingdom, by the ATCC and by the recently established Network on Antimicrobial Resi...

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Cloning and expression in Escherichia coli and Staphylococcus aureus of the beta-lysin determinant from Staphylococcus aureus: evidence that bacteriophage conversion of beta-lysin activity is caused by insertional inactivation of the beta-lysin determinant

Cited by 76 publications

References 42 publications

Enterotoxin production by Staphylococcus aureus isolates from cases of septicaemia and from healthy carriers

Enterotoxin production by Staphylococcus aureus isolates from cases of septicaemia and from healthy carriers

Molecular Cloning and Expression of Mn ²⁺ -Dependent Sphingomyelinase/Hemolysin of an Aquatic Bacterium, Pseudomonas sp. Strain TK4

Repair of Global Regulators inStaphylococcus aureus8325 and Comparative Analysis with Other Clinical Isolates

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Cloning and expression in Escherichia coli and Staphylococcus aureus of the beta-lysin determinant from Staphylococcus aureus: evidence that bacteriophage conversion of beta-lysin activity is caused by insertional inactivation of the beta-lysin determinant

Cited by 76 publications

References 42 publications

Enterotoxin production by Staphylococcus aureus isolates from cases of septicaemia and from healthy carriers

Enterotoxin production by Staphylococcus aureus isolates from cases of septicaemia and from healthy carriers

Molecular Cloning and Expression of Mn 2+ -Dependent Sphingomyelinase/Hemolysin of an Aquatic Bacterium, Pseudomonas sp. Strain TK4

Repair of Global Regulators inStaphylococcus aureus8325 and Comparative Analysis with Other Clinical Isolates

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Molecular Cloning and Expression of Mn ²⁺ -Dependent Sphingomyelinase/Hemolysin of an Aquatic Bacterium, Pseudomonas sp. Strain TK4