Resistance of messenger RNA-bound ribosomes to proteolytic dissociation.
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Biomedical subjects
Publications and source records attributed to M L Freedman.
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Cessation of exponential growth of Micrococcus radiodurans heightens X-radioresistance while either increasing or decreasing the deoxyribonucleic acid-protein ratio.
Short-term labeling with radioactive thymidine led to inaccurate estimates of deoxyribonucleic acid synthesis at some growth rates in steady-state cultures of Escherichia coli B/r. Estimates were corrected in chemostat cultures by adding adenosine, a known inhibitor of thymidine phosphorylase.
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Criteria are presented for distinguishing between synchronous and synchronized cultures (natural vs. forced synchrony) on the basis of characteristics of growth and division during a single generation. These criteria were applied in an examination of the uptake of potassium during the cell growth and division cycle in synchronous cultures and in a synchronized culture of Escherichia coli. In the synchronous cultures the uptake of (42)K doubled synchronously with cell number, corresponding to a constant rate of uptake per cell throughout the cell cycle. In the synchronized culture, uptake rates also remained constant during most of the cycle, but rates doubled abruptly well within the cycle. This constancy of (42)K uptake per cell supports an earlier interpretation for steady-state cultures that uptake is limited in each cell by a constant number of functional sites for binding, transport, or accumulation of compounds from the growth medium, and that the average number of such sites doubles late in each cell cycle. The abrupt doubling of the rate of uptake of potassium per cell in the synchronized culture appears because of partial uncoupling of cell division from activation or synthesis of these uptake sites.
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The average amount of deoxyribonucleic acid (DNA) per cell was measured in steady-state cultures of Escherichia coli B/r grown at 37 C in glucose-limited chemostats or in batch cultures in the exponential growth phase as maintained with one of several carbon sources. Within experimental errors, DNA content was dependent only on growth rate and independent of the type of culture, the carbon source, or the addition of growth factors. The amount of DNA per cell increased continuously with growth rate over the range of 0.02 to 3 divisions per hour. The data over the entire range of growth rates are in agreement with a constant time for a single replication point to traverse the entire genome, 47 min, and with cell division following 25 min after termination of replication. The measured amount of DNA per genome was 4.2 x 10(-15) g (or 2.5 x 10(9) daltons).
Escherichia coli B/r and B(s-1) ceased division and increased in mean cell volume soon after infection with T-even phage. The effect was obtained with wild-type or rapid lysis mutants, as well as with ultraviolet light-killed phage and with bacteriophage ghosts which lack deoxyribonucleic acid. The cell response did not require the presence of phage genetic material or the production of progeny phage. A Poisson distribution of the fraction of adsorbed phage at different multiplicites of infection indicates that one phage per bacterium will produce maximum increase in cell volume. T-even phage-resistant E. coli mutants showed no enlargement response, and phage T1, T3, and T7 elicited neither abrupt termination of cell division nor host cell enlargement. Infection with baseplate-defective T4D 12(-) amber mutants, which bind reversibly to but do not penetrate the bacterium, also had no effect. In vitro restoration of normal baseplate function in these defective viruses allowed phage adsorption and penetration and caused host cell division arrest and enlargement. These findings indicate that arrest of division and increase in mean cell volume occur together when a sensitive strain of E. coli is infected with T-even phage that adsorb and penetrate normally.
Division of Escherichia coli was stopped and mean cellular volume was increased after infection with T-even phage. This host cell enlargement was temperature-dependent, cyanide-sensitive, and stable in the presence of hypertonic medium. Enlargement ceased at about the same time that energy metabolism ceased. Initially, enlargement was accompanied by a decrease in mean cell density. Tritiated 2, 6-diaminopimelic acid was accumulated and incorporated into cold acid-insoluble material at the preinfection rate. These findings suggest that the effect on host cell size is only in part an osmotic phenomenon and that it also reflects continued growth of the surface of the infected cell in the absence of cell division.
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