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Biomedical subjects

A K Vidaver

Publications and source records attributed to A K Vidaver.

33 records · Page 2Linked to original sources

Typing of fluorescent phytopathogenic pseudomonads by bacteriocin production.

Several phytopathogenic fluorescent Pseudomonas species, primarily P. syringae, could be grouped into 16 bacteriocin producer groups, including new and previously described groups. At least 86% of the P. syringae strains could be typed by bacteriocin production. There was poor correlation between bacteriocin type and host plant origin. No correlation was detected between syringomycin (a phytotoxin) production and bacteriocin type.

Bacterial Toxins↗

Ultrastructure of bacteriophage phi 6: arrangement of the double-stranded RNA and envelope.

Pseudomonas phaseolicola cells synchronously infected with phi 6 were fixed and embedded by several procedures. The diameter of phi 6 measured 75 nm and its nucleocapsid 60 nm. Virus nucleocapsids were icosahedral and surrounded by a double membrane. In planar sections the nucleic acid appeared as a hexagonal ring and in cross section as two angular structures.

Bacteriophages↗

Synthesis of bacteriophage phi6 double-stranded ribonucleic acid.

Uracil was incorporated into all three bacteriophage phi6 dsRNA segments throughout the infection cycle; the rates of incorporation into each of the three segments were approx. constant for the first 15 to 20 min and then increased rapidly until 50 min after infection. The medium and small dsRNA segments were produced in greater amounts than the large dsRNA segment at all times in the infection cycle. Inhibition of host RNA and protein synthesis with rifampin and chloramphenicol revealed that virus dsRNA synthesis immediately after infection was independent of either host function.

Bacterial Proteins↗

Intermediates in the biosynthesis of double-stranded ribonucleic acids of bacteriophage phi 6.

Pseudomonas phaseolicola infected with bacteriophage phi 6 synthesized all three viral double-stranded RNA segments, three single-stranded RNAs, and three replicative intermediate-like RNAs in the presence of rifampin. The single-stranded RNA intermediates sedimented and electrophoresed along with melted viral double-stranded RNA, annealed with melted viral double-stranded RNA, and were transient in nature. The relative amounts of the single-stranded RNA intermediates varied during the infection cycle and were altered in the presence of chloramphenicol. The replicative intermediate-like RNAs sedimented faster than double-stranded RNA, failed to enter 2.5% polyacrylamide gels, eluted with double-stranded RNA from a CF-11 cellulose column, were precipitated with single-stranded RNA in 2 M LiC1, and yielded three genome-size pieces of double-stranded RNA upon digestion with RNase. These results are consistent with the hypothesis that complementary strands of the phi 6 double-stranded RNAs are synthesized asynchronously during the infection cycle.

Bacteriophages↗

Purification and characterization of syringacin 4-A, a bacteriocin from pseudomonas syringae 4-A.

Syringacin 4-A, a bacteriocin produced by Pseudomonas syrinagae 4-A, was obtained by induction with ultraviolet irradiation or mitomycin C. Approximately 1,000-fold purification of the bacteriocin was achieved by manganous chloride precipitation, differential centrifugation, and chromatography on hydroxyapatite columns. The purified syngacin was homogeneous on hydroxyapatite columns and sucrose density gradients; it also sedimented as a single entity in the analytical ultracentrifuge. The buoyant density of purified syringacin in cesium chloride was 1.294 g/ml. The sedimentation coefficient was calculated as 120S, and the diffusion coefficient was 6.49 x 10(-8) cm(2)/s. The molecular weight was calculated as 1.6 x 10(7) from physical data and 1.7 x 10(7) from biological data. The syringacin was composed of about 88.4% protein, 8.5% arabinose, 2.2% galacturonic acid, and 0.7% glucosamine. Amino acid analysis indicated a predominance of leucine (12.1%), aspartic acid (12.2%), and glutamic acid (12.7%). The ultraviolet spectrum showed a maximum absorbance peak at 276 nm. The syringacin was heat and alcohol sensitive, but resistant to trypsin, chymotrypsin, carboxypeptidase, Pronase, protease, lysozyme, steapsin, deoxyribonuclease, and ribonuclease. Maximum pH stability was between 5 and 8. Crude bacteriocin was stable at room temperature for at least a year, and purified material was stable for at least 3 months at 4 C.

Bacteriocins↗

Base composition and hybridization studies of the three double-stranded RNA segments of bacteriophage phi 6.

The three double-stranded ribonucleic acid (dsRNA) segments of the bacteriophage phi6 were isolated and shown to have similar melting temperatures and base compositions. RNA:RNA hybridization experiments with the isolated segments eliminate the possibility that the two smaller dsRNA segments arise from a cleavage of the large dsRNA segment. The two smaller RNA segments reanneal rapidly even at low temperatures; in contrast, the large dsRNA reannealed only at higher temperatures. Evidence is also presented which suggests that the dsRNAs may contain a short single-stranded RNA tail.

Bacteriophages↗

Bacteriophage phi6: a Lipid-Containing Virus of Pseudomonas phaseolicola.

The purification and properties of a lipid-containing bacteriophage, phi6, are described. The phage contains a lipid envelope which is probably essential for infection. Infectivity of phi6 was lost in the presence of organic solvents, sodium deoxycholate, and phospholipase A. The fatty acid composition of the phage lipid was similar to that of the Pseudomonas phaseolicola host cells. The phage was composed of about 25% lipid, 13% RNA, and 62% protein. The buoyant density of phi6 was 1.27 g/ml in cesium chloride. The morphology of phi6 was unusual; it had a polyhedral head of about 60 nm surrounded by a membranous, compressible envelope which appeared to assume an elongated configuration upon attachment to pili. The adsorption rate constant was 3.3 x 10(-10) ml/min in a semi-synthetic medium and 3.8 x 10(-10) ml/min in a nutrient broth-yeast extract medium. The latent period was shorter in the former medium (80-115 min compared with 120-160 min), and the average burst size was larger (250-400 compared with 125-150). The eclipse period coincided with the latent period.

Journal Article↗

RNA polymerase activity associated with bacteriophage phi 6.

The Pseudomonas phaseolicola bacteriophage phi6 incorporated labeled UTP into an acid-insoluble precipitate. Incorporation was dependent on the presence of manganese acetate, ATP, GTP, CTP, and a short heat treatment of the phage; the reaction was stimulated by NH(4)Cl. The substitution of (14)C-ATP, -CTP or -GTP for UTP, together with the appropriate unlabeled ribonucleoside triphosphates, disclosed that CMP was incorporated to the greatest extent followed by GMP, UMP, and AMP. Radioactive RNAs formed by the reaction were resistant to RNases A and T(1) in high salt but susceptible to these nucleases in low salt. The labeled RNA co-sedimented and co-electrophoresed with phi6 double-stranded (ds) RNA. However, the distribution of the radioactivity into the three ds-RNA components varied depending on the (14)C-ribonucleoside triphosphate used in the reaction. The incorporation of UMP was primarily into the two smaller ds-RNA segments, GMP primarily into the large ds-RNA segment, and CMP and AMP were about equally distributed into all three ds-RNA segments.

Acetates↗

Characterization of Xanthomonas phaseoli Bacteriophages.

Ten bacteriophages for Xanthomonas phaseoli were characterized. On the basis of adsorption rates, latent period and burst size, plaque morphology, host range, efficiency of plating, ultrastructure, sensitivity to osmotic shock, streptomycin sensitivity, temperature effects on plating efficiency, and serology, the phages were separated into at least three groups. Some of the phages were infectious for Pseudomonas phaseolicola (four strains) and P. syringae (one strain) in a narrow temperature range. The taxonomic and ecological significance of this finding is discussed.

Journal Article↗