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C I Kado

Publications and source records attributed to C I Kado.

At least 109 records · Page 6Linked to original sources
PubMed1978

Proteins conferred by the virulence-specifying plasmid of Agrobacterium tumefaciens C-58.

Membrane-associated and periplasmic proteins of Agrobacterium tumefaciens C-58 were compared with those from avirulent (nontumrigenic) derivative strains by slab and two-dimensional gel electrophoresis. Two proteins (Per-I and Per-2), with a molecular weight of 37,500 and 37,300, respectively, were detected in the supernatant fraction of cells of strain C-58 treated with EDTA and lysozyme in which a 117-megadalton plasmid confers virulence on the organism. The same proteins are missing in an avirulent plasmid-free derivative of C-58. When this derivative is mated with C-58, the resulting transconjugants regain the large C-58 plasmid together with the restoration of virulence and the expression of Per-1 and Per-2 proteins. When the transconjugants were cured of their plasmid, they concomitantly lost their virulence and Per-1 and Per-2. The functional roles of these proteins are unknown, but they are associated with the outer membrane and periplasmic fraction of the Agrobacterium cell. If directly involved in tumorigenesis, these proteins are not the sole determinants of tumorigenicity because they are synthesized in an avirulent derivative of C-58 that carries a deletion in the plasmid in the region conferring the tumorigenic phenotype. These results strongly suggest that the Per-1 and Per-2 proteins are plasmid-coded gene products. The possible roles of these proteins in specifying host range and host-cell attachment are also discussed.

Bacterial Proteins↗
PubMed1978

DNA modifying enzymes of Agrobacterium tumefaciens: effect of DNA topoisomerase, restriction endonuclease, and unique DNA endonuclease on plasmid and plant DNA.

Extracts from Agrobacterium tumefaciens strain ID135 contain three enzymes that have been characterized and partially purified. The first enzyme, a DNA topoisomerase, appeared to relax only negatively twisted DNA. The second enzyme, Atu I, a type II restriction endonuclease, generated the identical DNA digestion pattern as EcoRII when several DNAs were used. The third enzyme, endonuclease A, showed a preference for superhelical DNAs as substrates. When plasmid pCK135DNA, obtained from the virulent strain IDI135 of A. tumefaciens, or plant DNA was exposed to the three enzymes, changes in DNA patterns were observed due to either conformational changes or digestion of the DNAs. These enzymes may function in vivo in the processing and incorporation of bacterial DNA in plant cells.

Animals↗
PubMed1977

Escherichia coli plasmid pBR313 insertion into plant protoplasts and into their nuclei.

Cowpea mesophyll protoplasts were shown to bind irreversibly up to 3% input radioactive pBR313 plasmid DNA after 15 min of contact. Maximum uptake occurred in the presence of 5 mM ZnSO4 and 5 microgram/ml poly-L-ornithine. Under these conditions about one half of the TCA precipitable radioactivity was associated with the nuclear fraction and behaved as linear plasmid molecules. These could not be chased from the protoplasts upon further incubation with unlabeled plasmid DNA. The presence of donor DNA within the nuclear fraction is most probably not due to an artifactual redistribution of adsorbed plasmid DNA. Prolonged incubation periods resulted in extensive degradation of plasmid in the incubation medium but little degradation occurred in the protoplasts. The donor DNA was not covalently associated with the protoplast nuclear DNA.

Cell Nucleus↗
PubMed1977

Studies on Agrobacterium tumefaciens. VIII. Avirulence induced by temperature and ethidium bromide.

When tumorigenic strains of Agrobacterium tumefaciens were subcultured at temperatures between 31.5 and 37 degrees C or in broth containing ethidium bromide, they lost their capacity to induce tumors in tomato plants. The sensitivities of curing virulence (tumorigenicity) depended on the density of the population of cells, fewer cells (100/ml) being more sensitive to curing than higher densities (10(6)/ml). The loss of virulence need not require the total loss of the virulence-specifying plasmid, but may result from a loss of a small segment of that plasmid. Virulent strains made avirulent by temperature or ethidium bromide treatment still harbor a large plasmid of 70-80 megadaltons size compared with the 100- to 120-megadalton plasmid in the untreated strains.

Ethidium↗
PubMed1976

High molecular weight deoxyribonucleic acid polymerase from crown gall tumor cells of periwinkle (Vinca rosea).

A high molecular weight (6 S) plant DNA polymerase from axenic Vinca rosea tissue culture cells has been purified 2200-fold and characterized. The enzyme has a molecular weight of 105 000 (+/-5000). Sodium dodecyl sulfate-acrylamide gel electrophoresis of the purified enzyme yields polypeptide subunits having molecular weights of 70 000 and 34 000. The purified enzyme has a pH optimum of 7.5; a cation requirement optimum of 6 mM Mg2+ or 0.5 mM Mn2+; an apparent requirement for Zn2+; a Km of 1 muM for dTTP; and a 3.5-fold stimulation by 50 mM KCl. The enzyme is sensitive to N-ethylmaleimide (1 mM), heparin (0.1 muM), ethanol (5%), pyrophosphate (0.05 muM), and o-phenanthroline (0.1 mM) but is insensitive to rifamycin. Denatured DNA is found to be the best natural template, and only negligible activity can be demonstrated with the ribopolymer templates poly(dT)n-poly(rA)n and p(dT)10-poly(rA)n. In addition to the polymerization reaction, the enzyme catalyzes a pyrophosphate exchange reaction. Antibody to calf thymus 6-8S DNA polymerase does not inhibit DNA polymerase from Vinca rosea, suggesting no antigenic relationships between the mammalian and plant enzymes.

Binding, Competitive↗
PubMed1976

Polygalacturonic acid trans-eliminase in the osmotic shock fluid of Erwinia rubrifaciens: characterization of the purified enzyme and its effect on plant cells.

An endopolygalacturonic acid trans-eliminase (EC 4.2.2.2), released by osmotic shock of Erwinia rubrifaciens cells, has been purified to near homogeneity (3, 100-fold) by column chromatography on diethylaminoethyl-cellulose, phosphocellulose, and hydroxyapatite-cellulose followed by isoelectric focusing. It has a molecular weight of 41,000, s20,w of 3.09S, an isoelectric point of pH 6.25, pH optimum of 9.5, and a temperature optimum of 37 C and requires Ca2+ with an optimum concentration of 0.5 to 1.0 mM. Mg2+ could not substitute for Ca2+. Tyrosinyl residues seem essential for enzyme catalysis based on rapid inactivation by tetranitromethane. The enzyme prefers unmethylated polygalacturonic acid as the substrate, cleaving alpha-1,4-glycosidic linkages randomly to form unsaturated galacturonides at a Vmax of 1,166 mumol of product/min per mg of protein and a Km of 5 mg of polygalacturonic acid per ml. Over 90% of the enzyme activity is released from osmotically shocked E. rubrifaciens cells. Unlike E. rubrifaciens, trans-eliminase is not released from Erwinia carotovora cells by osmotic shock treatment, but enzyme activity is detected in the culture medium. The release of the enzyme is reduced fivefold by the addition of dibutyryl cyclic adenosine 5'-monophosphate. The hypersensitive reaction in tobacco leaves was induced within 60 min after injection of less than 1 mug of purified E. rubrifaciens trans-eliminase. Single cells of tobacco in suspension culture are readily killed by the enzyme, whereas tobacco protoplasts remain unaffected when treated in the same manner. These results indicate that endopolygalacturonic acid trans-eliminase is a constitutive enzyme possibly located in the periplasmic space of the E. rubrifaciens cell and releases enzyme into the culture medium in the presence of substrate. The release of the enzyme in tobacco tissue and the trans-eliminative cleavage of plant cell wall components may be steps leading to hypersensitivity of the tobacco tissue.

Calcium↗
PubMed1974

Pierce's disease of grapevines: evidence for a bacterial etiology.

A Gram-positive, rod-shaped bacterium was isolated and grown in pure culture on artificial mediums from the leafhopper Draeculacephala minerva Ball which had fed on plants infected with Pierce's disease. The bacterial culture was injected into uncontaminated leafhoppers which were than allowed to feed on healthy grapevines. After exposure to these leafhoppers, typical symptoms of Pierce's disease developed; however, no symptoms developed on plants exposed to leafhoppers injected with sterile culture medium. The same organism was reisolated from the experimentally inoculated plants. Electron microscopic examination of these infected plants revealed bacterial cells localized in the xylem tissues. No such cells were seen in healthy or control plants.

Bacteria↗
PubMed1974

Quantitative estimation of Agrobacterium tumefaciens DNA in crown gall tumor cells.

Several reports suggest that Agrobacterium tumefaciens nucleic acids can induce transformation of the cells of susceptible host plants and that bacteria-free tissue cultures of transformed cells contain A. tumefaciens DNA, RNA, antigens, or bacteriophages. We assayed Vinca rosea tumor DNA for base sequence homologies with A. tumefaciens DNA by DNA.DNA solution enrichment and DNA.DNA filter saturation hybridization techniques. No homologies were found by either method. The filter saturation hybridization technique included model filters containing known percentages of bacterial DNA mixed with V. rosea leaf DNA. Using this sensitive technique, we found that no more than 0.02% of the crown gall tumor genome could be homologous to A. tumefaciens DNA. This upper estimate of homology corresponds to 0.2 bacterial genome equivalent per diploid tumor cell.

Base Sequence↗