Determination of Genome Size of Selected Typing Bacteriophages of Staphylococcus aureus

The molecular weights of six representative typing bacteriophages were determined by sucrose gradient centrifugation. Genome size appeared to be related to the size of the phage head.     

Effect of Protein A on Adsorption of Bacteriophages to Staphylococcus aureus

Experiments were performed to determine if protein A influenced the association of bacteriophages with Staphylococcus aureus. Bacteriophage adsorption was compared in a S. aureus strain rich in protein A and mutants of this strain with very little protein A, in a strain with little protein A, and in mutants of this strain with increased protein A. In addition, the effect of growth in mannitol-salt broth and trypsin digestion (known to reduce protein A) on bacteriophage absorption was measured. There was an inverse relationship between protein A content of strains and the quantity of bacteriophage absorbed. However, no inhibition of staphylococcal phages was obtained with purified soluble protein A. Protein A as a surface component rendered the bacteria more resistant to adsorption of staphylococcal typing phages presumably by masking the phage receptor sites. When protein A-deficient mutants were incubated with bacteriophages, there was survival of staphylococci with increased protein A content probably due to a selective action.     

Base Ratio and Deoxyribonucleic Acid Homology Studies of Six Staphylococcus aureus Typing Bacteriophages

Genetic relatedness among Staphylococcus aureus typing bacteriophages 80, 47, 81, 71, 77, and 187 was investigated by using base ratio determinations and deoxyribonucleic acid (DNA)-DNA hybridization. Guanine/cytosine (G/C) content, as determined by thermal denaturation and chromatographic analysis of the purines released by acid hydrolysis of the DNA, was between 31 and 36%. No pattern correlating G/C content with serological or lytic group was discernible. DNA-DNA hybridization studies indicated high degrees of homology (43% or more) among the genomes of phages in the same serological group. Less homology (29% or less) was observed between the genomes of phages belonging to different serological groups. These findings implied a positive correlation between serological and genetic relatedness.     

Alpha-toxin of Staphylococcus aureus.

Alpha-toxin, the major cytotoxic agent elaborated by Staphylococcus aureus, was the first bacterial exotoxin to be identified as a pore former. The protein is secreted as a single-chain, water-soluble molecule of Mr 33,000. At low concentrations (less than 100 nM), the toxin binds to as yet unidentified, high-affinity acceptor sites that have been detected on a variety of cells including rabbit erythrocytes, human platelets, monocytes and endothelial cells. At high concentrations, the toxin additionally binds via nonspecific absorption to lipid bilayers; it can thus damage both cells lacking significant numbers of the acceptor and protein-free artificial lipid bilayers. Membrane damage occurs in both cases after membrane-bound toxin molecules collide via lateral diffusion to form ring-structured hexamers. The latter insert spontaneously into the lipid bilayer to form discrete transmembrane pores of effective diameter 1 to 2 nm. A hypothetical model is advanced in which the pore is lined by amphiphilic beta-sheets, one surface of which interacts with lipids whereas the other repels apolar membrane constitutents to force open an aqueous passage. The detrimental effects of alpha-toxin are due not only to the death of susceptible targets, but also to the presence of secondary cellular reactions that can be triggered via Ca2+ influx through the pores. Well-studied phenomena include the stimulation of arachidonic acid metabolism, triggering of granule exocytosis, and contractile dysfunction. Such processes cause profound long-range disturbances such as development of pulmonary edema and promotion of blood coagulation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Stress resistance in Staphylococcus aureus.

Staphylococcus aureus is a major human pathogen of increasing importance as a result of the spread of antibiotic resistance. It causes a wide range of diseases and survives outside the host by virtue of its adaptability and resistance to environmental stress. Several cellular components involved in Staphylococcus aureus stress resistance have begun to be characterized.     

Investigation of the Penicillinase Activity in L Colonies of Staphylococcus aureus

Penicillinase-producing strains of Staphylococcus aureus are transformed into stable L colonies by 70 to 100 subcultures on methicillin-containing medium with a suitable high osmolarity. During transformation, the penicillinase activity is lost. This loss in activity is not the result of only the penicillinase-negative mutants transforming to L colonies. If unstable L colonies are filtered through 0.45-mum membrane filters immediately after transformation, still no penicillinase activity is seen; this is also the case if the filtrated L colonies are reverted into coccal forms. The mechanism responsible for the loss of penicillinase activity is discussed. A loss of the penicillinase plasmid is proposed as the most reasonable explanation.     

Restriction-deficient mutants of Staphylococcus aureus.

A series of restriction-deficient mutants was isolated from non-lysogenic strains of Staphylococcus aureus belonging to phage groups I and II. Some mutants were sensitive to all phages tested. With one possible exception, all the mutants were unaffected in their modification systems. The breakdown of DNA of phages, restricted in the parental strains, was reduced in both the mutants that were tested. The restriction in propagating strain 3A could be transduced to its restriction-deficient mutant. The transduction efficiency increased after ultraviolet irradiation of the transducing phage suggesting that the gene for restriction is present on the bacterial chromosome.     

Prevention of Staphylococcus aureus lysis.

Staphylococcus aureus S-6 cells grown in chemically defined media often lysed after exponential growth. Lysis could be prevented by the addition of alanine or proline before the culture reached stationary phase.     

DNA replication in bacteriophage-infected Staphylococcus aureus.

The synthesis of viral and host DNA in phage-infected Staphylococcus aureus was examined. Three intracellular forms of phage 52HJD DNA were demonstrated: covalently closed circular, open circular, and linear DNA species. It was noted that infection of S. aureus-propagating strains 81 and 52 with phage 52HJD inhibited the replication of the bacterial chromosome and a stringently controlled penicillinase plasmid. A small tetracycline plasmid, normally under relaxed replication control, continued to replicate in the postinfection period. No breakdown of the host chromosome into small-molecular-weight fragments or utilization of bacterial DNA material for the synthesis of viral DNA was observed.     

Staphylococcus aureus in rural drinking water.

Coagulase-positive Staphylococcus aureus was isolated from over 6% of 320 rural drinking water specimens. Well water was the most common source examined. The presence of S. aureus was not found to correlate with the presence of coliform bacteria. Strains of Staphylococcus that produced enterotoxin A were found in 40% of the samples containing S. aureus. Additional studies showed that faucet aerator screens were common sources of high cell densities of S. aureus.     

Plasmid-protein relaxation complexes in Staphylococcus aureus.

Protein-deoxyribonucleic acid relaxation complexes have been demonstrated for six Staphylococcus aureus plasmids out of sixteen examined. Four of these encode stretomycin resistence, have molecular weights of about 2.7 x 10(6), and are isolated as supercoiled molecules that are virtally 100% relaxable by treatment with sodium dodecyl sulfate. It is probable that these four isolates represent a single widely disseminated plasmid species. The other two plasmids showing relaxation complexes have molecular weights of about 3 x 10(6) and encode chloramphenicol resistance. The complexes in these cases are unstable, and it has not been possible to induce more than 50% relaxation by any of the standard treatments. Ten other plasmids do not show detectable complexes. These include three penicillinase plasmids, four tetracycline-resistance plasmids, one plasmid carrying kanamycin-neomycin resistance, and finally, two chloramphenicol-resistance plasmids.