Glucosylation of Teichoic Acid: Solubilization and Partial Characterization of the Uridine Diphosphoglucose:Polyglycerolteichoic Acid Glucosyl Transferase from Membranes of Bacillus subtilis

Polyglycerolteichoic acid:glucosyl transferase (TAG transferase), one of the three enzymes involved in the pathway leading to the glucosylation of teichoic acid in Bacillus subtilis 168, was investigated. During the early stages of the growth of B. subtilis, TAG transferase is predominantly a soluble enzyme found in the cytoplasm. As growth proceeds, the amount of soluble enzyme decreases and the proportion of insoluble, membrane-bound TAG transferase increases, reaching a maximal value at the close of the logarithmic phase. Data are presented which suggest that these are two forms of the same enzyme, or have some common component. The effects of chaotropic agents, such as sodium trichloroacetate and sodium perchlorate, on the cytoplasmic membrane were also studied. These data show that such compounds can effectively remove the TAG transferase from the membrane in a water-soluble form. A study of some of the physical properties of this solubilized enzyme suggests that there is little difference between the two forms of the enzyme. Experiments are described which indicate that the glucosyl transfer by both the membrane-bound and soluble enzymes is not mediated by lipids.     

Salt injury to plants with special reference to cations versus anions and ion activities

Summary In contrast with the toxicities of sulfate and chloride salts added to substrates, the anions SO₄ and C1 were not injurious when accumulated without leaf burning by cotton and tomato plants from atmospheres enriched with SO₂ or HC1 gases. The foregoing results are discussed in terms of cationenzyme interactions which appear to represent at least a major cause of salt toxicity. Although anions are largely unreactive with enzymes it has long been observed that chloride salts in soil solutions are far more toxic than sulfate salts. Five of seven species have shown nearly equal growth repressions on substrates with 100 me/1 of C1 salts versus 200 me of SO₄ salts, each added as 50 per cent Na. The ion activities of the two solutions were equal and the sum of cations in the plant saps were similar. The osmotic differentials (average about 10 atm) between the expressed tissue fluids and these substrate solutions were remarkably uniform within species. It is projected that the downward transport of salts via the phloem provides for root concentrations which supply ions to the xylem and thereby control the uptake of substrate salts.     

Inhibition of Transformation of Bacillus subtilis by Heavy Metals

Mercuric ions, as well as organomercuric ions and cadmium ions, can inhibit deoxyribonucleic acid-mediated transformation in Bacillus subtilis 168 without decreasing the viability of the total population. Differences in the inhibition of transformation by mercuric ions are identifiable on a temporal and concentration dependence basis. Sensitivity to low concentrations (9.2 x 10(-8) M) appears early in the uptake of deoxyribonucleic acid before the transformed markers have become insensitive to deoxyribonuclease. Resistance to "low concentrations" of Hg(2+) is kinetically indistinguishable from the requirement for magnesium in the transformation process. This inactivation is not reversed by the mercury-binding compound glutathione. Sensitivity to mercuric ions at a higher concentration (5.52 x 10(-7) M) occurs after the donor deoxyribonucleic acid has become insensitive to deoxyribonuclease. These complex interactions between mercuric ions and the process of transformation are discussed.     

Mutation in Saccharomyces cerevisiae Conferring Streptomycin and Cold Sensitivity by Affecting Ribosome Formation and Function

A cold-sensitive, streptomycin-sensitive mutant of Saccharomyces cerevisiae accumulates a 28S ribonucleoprotein particle when grown at low temperature. This particle contains 17S ribosomal ribonculeic acid which is degraded when exposed to ribonuclease. The particle does not serve as a precursor to 60 and 40S ribosomal subunits nor is it turned over when growth is allowed to resume at the permissive temperature; rather it is only diluted by growth. That streptomycin sensitivity (allelic with cold sensitivity) is ribosomal is evidenced by the inhibition of protein synthesis in vitro by streptomycin and the binding of labeled streptomycin to the mutant but not the parental 40S ribosomal subunit.     

Conformational aspect of polypeptide structure. XLIV. Conformational transitions of poly(N-methyl-alanines) induced by trifluoroacetic acid

The synthesis of poly(N-methyl-L -alanine) and poly (N-methyl-DL -alanine) are described. The polymers were examined by 220 MHz high-resolution nuclear magnetic resonance (nmr) and circular dichroism (CD). The results demonstrate that poly(N-methyl-L -alanine) exists as an ordered helical structure with all the amide bonds in the trans configuration in appropriate solvents. As trifluoroacetic acid (TFA) is added to the solutions of the polymer in helix-supporting solvents, resonances corresponding to both trans and cis amide conformations of N-methyl, C-methyl, and α-CH are observed. The presence of both the trans and the cis peptide bonds in a polymer chain disrupts the ordered structures. Our conclusions from CD data are in agreement with the nmr results. Ultracentrifugation shows that degradation of the polymer chain does not occur during the TFA treatment.     

The macronuclear envelope of Tetrahymena pyriformis GL in different physiological states

Summary Macronuclear envelopes were isolated from the ciliated protozoan Tetrahymena pyriformis GL, negatively stained and examined in the electron microscope. The frequency of central granules in the macronuclear pores was evaluated in five different physiological states: (1) stationary phase of growth, (2) exponential phase of growth, (3) heat-synchronized cultures at the end of the heat-synchronization treatment, (4) heat-synchronized cultures at the beginning of the first division, (5) heat-synchronized cultures at the end of the first division.The percentage of pores containing a central granule was markedly enhanced in heatsynchronized cultures at the end of the first division, i.e. a state known for an increase in ribosome formation. Actinomycin D was found to cause a significant decrease in central granule frequency.The observed alterations in central granule frequency seem to confirm the hypotheses which consider the central granule as representing a ribonucleoprotein particle in transit from nucleus to cytoplasm through the nuclear pore.For careful technical assistance I am indebted to Miss Marianne Whiter as well as to Drs. H. Falk, W.W. Franke and P. Sitte for helpful discussions. This work was supported in part by the Deutsche Forschungsgemeinschaft.