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L Mindich

Publications and source records attributed to L Mindich.

70 records · Page 4Linked to original sources

Isolation of nonsense suppressor mutants in Pseudomonas.

A strain of Escherichia coli harboring the drug resistance plasmid RP1 was treated with the mutagen N-methyl-N-nitro-N-nitro-N-nitrosoguanidine, and mutants were isolated in which ampicillin resistance had been lost due to an amber mutation in the plasmid. One of these mutants was again treated, and a strain was isolated in which tetracycline resistance was also lost due to an amber mutation in the plasmid. The plasmid containing amber mutations in the genes amp and tet was named pLM2. This plasmid could be transferred to strains of Pseudomonas aeruginosa, P. phaseolicola, and P. pseudoalcaligenes. Mutants resistant to ampicillin and tetracycline could not be obtained from P. phaseolicola carrying pLM2. However, strains of E. coli, P. aeruginosa, and P. pseudoalcaligenes carrying the plasmid did produce mutants simultaneously resistant to both antibiotics. All of the mutants of E. coli had developed nonsense suppressors since they became phenotypically lac+, although harboring a lac amber mutation, and formed plaques with amber mutants of phages PRR1 and PRD1 that attack organisms carrying RP1. Approximately 20% of the resistant mutants of P. aeruginosa and P. pseudoalcaligenes were sensitive to the amber mutant of PRD1. These mutants were of variable stability and grew somewhat more slowly than their parent strains. One of the suppressor mutants of P. pseudoalcaligenes, designated ERA(pLM2)S4, was used for the isolation of nonsense mutants of bacteriophage PHA6, a virus having a segmented genome of double-stranded ribonucleic acid and an envelope of lipids and proteins.

Ampicillin↗

Isolation and characterization of a glycerol auxotroph of Rhodopseudomonas capsulata: effect of lipid synthesis on the synthesis of photosynthetic pigments.

A glycerol auxotroph was isolated from Rhodopseudomonas capsulata for use as a system for studying membrane synthesis and function. When the mutant was deprived of glycerol, net phospholipid synthesis ceased immediately and a small amount of free fatty acids accumulated. A turnover of lipid occurred in both deprived and supplemented cultures. Deoxyribonucleic acid and protein synthesis continued for one doubling of cell massand then slowed down in deprived cells. Net ribonucleic acid synthesis slowed down more dramatically. Oxidative phosphorylation activity of membrane preparations from aerobically and semi-anaerobically grown cells appeared unaffected by glycerol deprivation, indicating that simultaneous lipid synthesis is not a requirement for new oxidative phosphorylating activity. In the absence of net phospholipid synthesis, bacteriochlorophyll and carotenoid syntheses were reduced to 30% of the activity of supplemented cultures. Delta-Aminolevulinic acid synthase, the first enzyme on the bacteriochlorophyll pathway that is subject to regulatory control, increased in activity in deprived cultures. Lascelles and Szilagyi (1965) showed an association between phospholipid synthesis and pigment production. They found an increased lipid content associated with pigmented cells. The present results indicate that not only is there an association between lipid and pigment synthesis, but also there is actually a dependence of bacteriochlorophyll synthesis on phospholipid synthesis.

5-Aminolevulinate Synthetase↗

Proteins of bacteriophage phi6.

We investigated the protein composition of the lipid-containing bacteriophage phi 6. We also studied the synthesis of phage-specific proteins in the host bacterium Pseudomonas phaseolicola HB10Y. The virion was found to contain 10 proteins of the following molecular weights: P1, 93,000; P2, 88,000; P3, 84,000; P4, 36,800; P5, 24,000; P6, 21,000; P7, 19,900; P8, 10,500; P9, 8,700; and P10, less than 6,000. Proteins P3, P9, and P10 were completely extracted from the virion with 1% Triton X-100. Protein P6 was partially extracted. Proteins P8 and P9 were purified by column chromatography. The amino acid composition of P9 was determined and was found to lack methionine. Labeling of viral proteins with [35S]methionine in infected cells indicated that proteins P5, P9, P10, and P11 lacked methionine. Treatment of host cells with UV light before infection allowed the synthesis of P1, P2, P4, and P7; however, the extent of viral protein synthesis fell off exponentially with increasing delay time between irradiation and infection. Treatment of host cells with rifampin during infection allowed preferential synthesis of viral proteins, but the extent of synthesis also fell off exponentially with increasing delay time between the addition of rifampin and the addition of radioactive amino acids. All of the virion proteins were seen in gels prepared from rifampin-treated infected cells. In addition, two proteins, P11 and P12, were observed; their molecular weights were 25,200 and 20,100, respectively. Proteins P1, P2, P4, and P7 were synthesized early, whereas the rest began to increase at 45 min post-infection.

Amino Acids↗

A mutant of Escherichia coli defective in DNA polymerase II activity.

A mutant of E. coli defective in DNA polymerase II activity was isolated. Extracts had about 0.1% of the normal activity of the DNA polymerase. The effect of the mutation on the ability of cells to replicate DNA of various sources was analyzed. Mutant bacteria grow normally, at 41 degrees as well as at 30 degrees . All bacteriophages, F factors, and R factors examined so far grow normally in the mutant. Sensitivity of the mutant to ultraviolet radiation and alkylating reagents in growth media was the same as that of the wild type. The mutation, polB1, is recessive with respect to wild type. The polB1 mutation can coexist with recombination-defective mutations. Genetic mapping studies show the mutation to be located at about 2 min on the E. coli map.

Chromatography↗

Membrane synthesis in Bacillus subtilis. 3. The morphological localization of the sites of membrane synthesis.

The results of several lines of investigation indicate that membrane growth in Bacillus subtilis does not occur at one or a small number of discrete zones. No indications of large regions of membrane conservation were observed. Kinetic labeling experiments of mesosomal and plasma membrane lipids indicate that the mesosomal lipids are not precursors of the plasma membrane lipids. Density shift experiments, in which the changes in buoyant density of membranes were studied after growth in deuterated media, showed no indication of large zones of conservation during membrane growth. Radioautography of thin sections of cells pulse labeled with tritiated glycerol showed no indication of specific zones of lipid synthesis. The consequences of these results for models of cell growth and division are discussed.

Autoradiography↗

Control of fatty acid synthesis in bacteria.

When glycerol-requiring auxotrophs of Bacillus subtilis are deprived of glycerol, the synthesis of fatty acids continues at an apparent rate of 20 to 50% that of supplemented cultures. The newly synthesized fatty acids are not incorporated into phospholipid and accumulate as free fatty acids. These molecules undergo a much more rapid turnover than phospholipid fatty acids, and the rate of turnover is sufficient to indicate that the rate of fatty acid synthesis in glycerol-deprived cultures is similar to that in supplemented ones. The average chain length of the free fatty acids is greater than that of the phospholipid fatty acids. Cells deprived of required amino acids also show a diminution in the apparent rate of fatty acid synthesis; however, in this case, the fatty acids accumulate in phospholipid, and no increase of the free fatty acid fraction is observed. It is argued on the basis of these findings that the control of lipid synthesis does not operate at the level of transacylation but must act on one or more of the reactions of the fatty acid synthetase.

Acetates↗

Induction of Staphylococcus aureus Lactose Permease in the Absence of Glycerolipid Synthesis.

Glycerol auxotrophs of S. aureus were isolated and shown to cease phospholipid synthesis immediately when deprived of glycerol. Second-step mutants with temperature-sensitive inducibility of the lac system were also isolated. When cells were induced by temperature shift to produce the products of the lac system in the absence of glycerol, the permease activity, relative to 6-phospho-beta-galactosidase activity, was between 30 and 50% that of glycerol-supplemented cultures. However, the phosphotransferase activity for beta-galactosides in isolated membranes was found to be normal when compared to the level of beta-galactosidase. This indicated that the permeation system was induced and integrated into the membrane, but did not function efficiently for transport. Readdition of glycerol in the presence of chloramphenicol resulted in a slow increase in efficiency of the transport activity. Glycerol deprivation after induction led to a small loss of permease efficiency.

Journal Article↗

Induction of citrate transport in Bacillus subtilis during the absence of phospholipid synthesis.

Citrate transport can be induced in a glycerol-requiring mutant of Bacillus subtilis even after deprival of glycerol. Under these conditions de novo phospholipid synthesis is stopped although ribonucleic acid, deoxyribonucleic acid, and protein syntheses continue. It is concluded that the inducibility of bacterial transport systems must not necessarily require de novo phospholipid synthesis.

Acetates↗

Bacteriocins of Diplococcus pneumoniae. I. Antagonistic relationships and genetic transformations.

Mindich, Leonard (The Public Health Research Institute of the City of New York, Inc., New York, N.Y.). Bacteriocins of Diplococcus pneumoniae. I. Antagonistic relationships and genetic transformations. J. Bacteriol. 92:1090-1098. 1966.-Several strains of Diplococcus pneumoniae produced bacteriocins inhibiting the growth of other strains of pneumococcus but inactive upon themselves. Strain R6, which did not produce pneumocin under most conditions, was changed to a producing state by mutation or by transformation with deoxyribonucleic acid from other pneumococci. The genetic determinants of several different bacteriocins were introduced into the same cell, where they acted independently. Determinants active in bacteriocin production were also constructed by crosses involving two nonproducing parents. Resistance to pneumocin B4 was a property of cells of the P4 type which produced it. On selection of pneumocin-resistant cells resulting from mutation or transformation in strain R6, resistant cells, all of which were pneumocin producers, were obtained. The genetic determinants of B4 production and resistance were transferred together in transformation. A gene for high level B4 resistance was found in strain P7S. This gene was not associated with B4 production and did not induce production when introduced into R6, although it did increase resistance to B4. Pneumocin B4 was bactericidal and caused a rapid cessation of growth. Its action was inhibited by pronase, trypsin, chymotrypsin, and phospholipids.

Anti-Bacterial Agents↗