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Biomedical subjects

D Schlessinger

Publications and source records attributed to D Schlessinger.

At least 235 records · Page 13Linked to original sources
PubMed1974

Characterization of the binding of amphotericin B to Saccharomyces cerevisiae and relationship to the antifungal effects.

Based on the enhanced fluorescence of amphotericin B in acid solutions, a quantitative assay for this polyene antibiotic has been developed that is sensitive and linear in the range of 0.1 to 10.0 muM. The binding of amphotericin B to Saccharomyces cerevisiae was assayed under various conditions as the amount bound to cells in a dialysis chamber or after centrifugation. Two types of binding were defined: weak, reversible binding occurred at 0 C or higher temperatures and even in the presence of inhibitors of energy metabolism, whereas strong, irreversible binding did not occur at 0 C and was inhibited when energy metabolism was blocked. Only strong binding was correlated with cell killing. Weak binding probably involves the outer layer of the membrane; strong binding probably requires disruption of hydrophobic regions of the cell membrane.

Amphotericin B↗
PubMed1974

Magic spot metabolism in an Escherichia coli mutant temperature sensitive in elongation factor Ts.

A temperature-sensitive mutant of Escherichia coli HAK88 which has been shown to have a lesion in elongation factor Ts (EFTs) was studied with repsect to its metabolism of guanosine 5'-diphosphate, 2'(3')-diphosphate (ppGpp) and the associated failure of ribosomal ribonucleic acid (rRNA) accumulation at the nonpermissive temperature. Results reported here show that (i) when EFTs is nonfunctional, a full complement of charged transfer RNA (tRNA) cannot prevent accumulation of ppGpp (magic spot) and the stringent failure of rRNA accumulation; (ii) chloramphenicol prevents magic spot (MS) formation and the stringent response not by increasing the percentage of charged tRNA, but possibly by somehow interfering directly with the synthesis of MS; and (iii) tetracycline can lead to MS disappearance without resumption of RNA synthesis. Thus, the absence of MS and the presence of a functional RNA polymerase and charged tRNA are not sufficient to support rRNA accumulation in vivo. An additional element in the regulatory system is suggested.

Adenosine Triphosphate↗
PubMed1974

Ribonucleic acid polymerases of the yeast phase of Histoplasma capsulatum.

Ribonucleic acid (RNA) polymerases of Histoplasma capsulatum (yeast phase) were fractionated by phosphocellulose chromatography and partially characterized. Three distinct, active fractions were seen. The major RNA polymerase species was inhibited strongly by alpha-amanitin, whereas the other two were resistant. When either slightly purified (HSE) extract or the major active component was assayed at 37 C, the incorporation of tritiated uridine monophosphate into RNA stopped after 10 to 15 min. In contrast, the synthesis continued for at least 1 h at 23 C. The other two RNA polymerase species exhibited higher rates of incorporation when tested at 37 C, and continued to synthesize RNA even after 60 min. However, by that time the levels of incorporation at 23 C were higher than at 37 C for all three enzymes. The temperature sensitivity was not affected by changing substrate concentration or employing either native or denatured calf thymus deoxyribonucleic acid as a template. These results are compared with the data obtained with RNA polymerases from different fungi and other organisms. A possible involvement of RNA polymerase(s) in morphological differentiation of H. capsulatum is discussed.

Ammonium Sulfate↗
PubMed1974

Polypeptide formation and polyribosomes in Escherichia coli treated with chloramphenicol.

In Escherichia coli cultures maximally inhibited with chloramphenicol, formation of polypeptides still continued at a slow, constant rate for at least 90 min. The rate of leucine incorporation was reduced to 0.5%, but methionine was only reduced to 2%, suggesting that chains are normally initiated with methionine but are prematurely released at a short chain length. Consistent with this possibility was the distribution of the products on Sephadex columns: a range of peptides longer than 4 and shorter than 60 to 70 residues was seen. Less than 10% of the peptides broke down during a chase with cold amino acids, and during continuous labeling they accumulated progressively. On the average, one peptide was formed per ribosome every 5 min. Peptide synthesis in the presence of chloramphenicol was still dependent on ribosome translocation; it stopped in a mutant with an inactivated temperature-sensitive elongation factor G. But even in the absence of translocation, new messenger ribonucleic acid (mRNA) chains were found joined to one or a few ribosomes. The chains had a size distribution comparable to that of mRNA from polyribosomes of growing cells. They were stabilized for an average time of about 5 min, but were more rapidly degraded after puromycin was added to the cells. This suggests that stabilization may be related to the average time spent by a ribosome on an mRNA chain, with or without polypeptide formation.

Amino Acids↗
PubMed1973

Synthesis of a large precursor to ribosomal RNA in a mutant of Escherichia coli.

A mutant of E. coli, isolated by Kindler and Hofschneider as a strain defective in RNase III activity, forms a 30S precursor of ribosomal RNA ("30S pre-rRNA"). The half-life of the 30S pre-rRNA in growing cells at 30 degrees , estimated by the rate of specific (3)[H]uridine incorporation, is about 1 min. In rifampicin-treated cells, the RNA is metabolized to mature rRNA with a half-life of about 2 min. The 30S pre-rRNA has been highly purified. DNA-RNA hybridization tests demonstrate that it contains both 16S and 23S rRNA sequences. Also, in cultures treated with rifampicin, the cleavage products of radioactive 30S pre-rRNA include 25S and 17.5S RNA species, destined to becomes 23S and 16S rRNA. Thus, each 30S chain probably contains one 16S and one 23S RNA sequence, as well as additional sequences. Two independent techniques indicate that the additional portions account for about 27% of the total lenght: (1) By comparison to the sedimentation rate and electrophoretic mobility of marker RNAs, the 30S pre-RNA has an apparent molecular weight of 2.3 x 10(6) +/- 5%, or 28% more than the sum of 16S and 23S rRNA; (2) 27% of the 30S pre-rRNA is not competed away from hybridization by mature 16S and 23S rRNA.Thus, bacteria appear to make a pre-rRNA similar in some respects to that observed in eukaryotes; though in normal E. coli cells, the pre-rRNA is ordinarily cleaved endonucleolytically during its formation.

Carbon Isotopes↗
PubMed1972

Antifungal properties of polymyxin B and its potentiation of tetracycline as an antifungal agent.

High concentrations of polymyxin B inhibited the growth of Candida albicans and Saccharomyces cerevisiae. When these yeasts were incubated with concentrations of polymyxin B too low to affect growth, and were then exposed to tetracycline, protein synthesis was inhibited and at least 99% of the organisms were killed. Neither inhibition of protein synthesis nor cell death occurred in cultures treated with high concentrations of tetracycline alone. We conclude that polymyxin B at high concentrations affects the cell membrane of yeasts, which results in inhibition of growth. At low concentrations, it increases the permeability of the yeast cell membrane to tetracycline, which then inhibits protein synthesis and leads to cell death.

Antifungal Agents↗