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Biomedical subjects

M Schaechter

Publications and source records attributed to M Schaechter.

30 records · Page 2Linked to original sources

Effect of rifampin on the structure and membrane attachment of the nucleoid of Escherichia coli.

Deoxyribonucleic acid (DNA) of Escherichia coli was found to be attached to the cell membrane at about 20 points. This was determined by fractionation of X-irradiated cells with the M band (magnesium-Sarkosyl crystals) technique. The number of attachment points was computed from the relationship between the amount of DNA in M bands and the number of double-strand breaks introduced by the X-ray treatment. The number of attachment points was decreased fourfold by treatment of cells with rifampin. This effect was apparently due to the action of the drug on ribonucleic acid (RNA) polymerase since the drug did not affect a mutant whose RNA polymerase is resistant to rifampin. This suggests that there may be two classes of attachment points of DNA on the membrane, some of which are removed by rifampin treatment and some which are not. Rifampin treatment also resulted in the uncondensing of isolated nucleoids and in an axial appearance of the nucleoids in ultrathin sections. The results suggest that RNA polymerase plays a role, direct or indirect, in maintaining the structure of the bacterial nucleoid and in some of its attachment to the membrane.

Carbon Radioisotopes↗

Bacteriophage T4 head maturation: release of progeny DNA from the host cell membrane.

We have presented a new approach to studying bacteriophage T4 head maturation. Using a modified M-band technique, we have shown that progeny deoxyribonucleic acid (DNA) was synthesized on the host cell membrane throughout infection. This DNA was released from the membrane later in infection as the result of formation of the phage head; detachment of the DNA required the action of gene products 20, 21, 22, 23, 24, 31, 16, 17 and 49, known to be necessary for normal head formation. Gene products 2, 4, 50, 64, 65, 13 and 14, also involved in head morphogenesis were not required to detach progeny DNA from the membrane; the presence of the phage tail and tail fibers also was not required. DNA was released in the form of immature heads and initially was sensitive to deoxyribonuclease (DNase). Conversion to DNase resistance followed rapidly. The amount of phage precursors present at the time of DNA synthesis determined the time of onset and detachment rate of DNA from the M band as well as the kinetics by which the detached DNA become DNase resistant.

Cell Membrane↗

The mode of segregation of the bacterial cell membrane.

The distribution of phospholipids from labeled parental membranes to progeny cells was studied by autoradiography and a minicell system. The minicell experiments showed that, during growth, the parental membrane is diluted at the same rate in cells and in minicells, which indicates that ends of cells are not different from the cylindrical portions with regard to the distribution of parental molecules. The same result was obtained after labeling heme-containing proteins with delta-aminolevulinic acid. The autoradiographic experiments indicate that the membrane segregates in about 250 subunits 4 x 10(4) nm(2) in size. These subunits appear to be conserved during growth.

Autoradiography↗

Some unique properties of the deoxyribonucleic acid-bearing portion of the bacterial membrane.

By using the M-band technique we have shown that portions of the membranes of Bacillus megaterium and Escherichia coli vary in their affinity for magnesium-Sarkosyl crystals and in phospholipid composition. The portion to which deoxyribonucleic acid is attached comprises as little as 4% of the total cell membrane, has a particularly high degree of affinity for magnesium-Sarkosyl crystals, and is rich in phosphatidylethanolamine. The M-band fractionation does not depend on the use of lysozyme.

Bacillus megaterium↗

Effect of shift-down and growth inhibition on phospholipid metabolism of Escherichia coli.

The metabolism of phospholipids of Escherichia coli was studied under conditions which inhibit various metabolic processes. Phospholipid synthesis and turnover were not inhibited by growth-inhibitory amounts of various antibiotics. Turnover of phosphatidylglycerol (PG) was inhibited by small amounts of dinitrophenol and by anaerobiosis. Turnover of phosphatidylethanolamine (PE), which is not detected in control cultures, was demonstrated under conditions of incipient lysis. When cells were shifted down from a rich to a poor medium, PE synthesis was inhibited, and incorporation of glycerol into the distal position of PG was stimulated. Under these conditions, turnover of the phosphate and the acylated glycerol moieties of PG was inhibited. Increased synthesis of PE was detected when filamentous cells were induced to make septa. The results indicate that PE synthesis is related to growth and cell division, whereas PG metabolism is related to other cell processes.

Anaerobiosis↗