1. The size distribution of aggregates of liver ribosomes and their protein-synthesizing ability in vitro were studied shortly after X-irradiation of guinea pigs. 2. Sucrose-density-gradient analysis of the mitochondrial supernatant after treatment with deoxycholate revealed a gradual increase in the number of polysomes, reaching a maximum between 9 and 15 hr. after irradiation. At that period the amount of ribonucleoprotein particles reached a level 25-30% above the control. This finding was confirmed by analytical-ultracentrifugal analysis and electron microscopy. Experiments were conducted to exclude the possibility that the enrichment of polysomes in the irradiated animals had occurred during the isolation procedure. 3. The protein-synthesizing ability of total ribosomal particles was measured in vitro. This showed an increase in amino acid incorporation parallel to the progressive enrichment of polysomes. At radiation doses of up to 1000r. the protein-synthesizing capacity was dependent on the radiation dose: the higher the dose the higher the amino acid incorporation, reaching 40-60% above the control at the period of maximal polysome enrichment. Amino acid incorporation remained at this level after radiation doses of between 1000 and 3000r. The enhanced protein-synthesizing activity was due solely to the increase in the proportion of polysomes, since irradiation was without effect on the activity of single ribosomes. 4. The results of the experiments are discussed in the light of our knowledge of the effect of radiation on protein synthesis.

Investigations have been carried out concerning the effects of microwave (MW) exposure on the aminoacyl‐transfer ribonucleic acid (tRNA) synthetase of the progeny of females that were exposed during their entire period of gestation (19 days). The changes caused by continuous‐wave (CW) and amplitude‐modulated (AM) MW radiation have been compared. CFLP mice were exposed to MW radiation for 100 min each day in an anechoic room. The MW frequency was 2.45 GHz, and the amplitude modulation had a 50 Hz rectangular waveform (on/off ratio, 50/50%). The average power density exposure was 3 mW/cm2, and the whole body specific absorption rate (SAR) was 4.23 ± 0.63 W/kg. The weight and mortality of the progeny were followed until postnatal day 24. Aminoacyl‐tRNA synthetase enzymes and tRNA from the brains and livers of the offspring (461 exposed, 487 control) were isolated. The aminoacyl‐tRNA synthetase activities were determined. The postnatal increase of body weight and organ weight was not influenced by the prenatal MW radiation. The activity of enzyme isolated from the brain showed a significant decrease after CW MW exposure, but the changes were not significant after 50 Hz AM MW exposure. The activity of the enzyme isolated from liver increased under CW and 50 Hz modulated MW. © 1996 Wiley‐Liss, Inc.

1. Liver RNA synthesis was studied within 24h after whole-body X-irradiation of guinea pigs that had been starved for 22-24h. 2. Microsomal RNA was labelled in vivo for 3h with [(14)C]orotic acid and the isolated labelled RNA was fractionated by sucrose-density-gradient centrifugation. Incorporation was 50-100% higher between 3 and 12h after 2000rd X-irradiation and at 22h was not elevated any further. Whole nuclear RNA was labelled with [(14)C]orotic acid for 15min. At 5h after irradiation the incorporation showed a 50-100% increase. Incorporation increased in all types of RNA studied. 3. The RNA phosphorus/DNA phosphorus ratio of whole liver gradually increased after X-irradiation. Maximal increase was found between 24 and 36h, which corresponds to a value about 40% above that of the starved control. The RNA phosphorus content of isolated ribonucleoproteins obtained from various cell fractions of the liver was similarly increased after X-irradiation. 4. Liver microsomes were obtained from X-irradiated and control animals. Microsomes were incubated in vitro with [(14)C]phenylalanine in the presence and absence of polyuridylic acid. After the incubation the microsomes were fractionated by sucrose-density-gradient centrifugation. The polyuridylic acid enhancement was twice as great in the microsomes of the control preparation as in the irradiated one. The experiment demonstrated a higher saturation of microsomes by endogenous messenger after X-irradiation. 5. RNA polymerase activity of the purified nuclear preparation was assayed. The activity of the Mg(2+)-dependent RNA polymerase activity was 50 and 200% respectively above the control values at 6 and 9h after X-irradiation. 6. Animals were treated with actinomycin D shortly before X-irradiation. This treatment abolished the radiation-induced enrichment of polyribosomes and the increase of protein-synthesizing activity. The effect of X-irradiation on the transcription of the genetic code of the liver is discussed.

The sensitivity in vitro against gamma-radiation of valyl-tRNA synthetase (VRS) from chick embryo brain proved to be higher than that of liver VRS. In order to study this phenomenon properties of VRS isolated from both organs were compared. Both enzymes have the same molecular weight (Mr = 110 000) determined by equilibrium sedimentation and by gel filtration. Electrophoresis on polyacrylamide gel in the presence of sodium dodecyl sulphate showed no evidence for subunit structure. The optimal reaction conditions in valyl-tRNA formation were found to be the same, except the pH values. Km values of the reactions were also similar. Both enzymes reacted with the tRNAs isolated from the other organ. The stability during storage of the liver VRS was higher than that of the brain VRS. In the brain VRS molecule one buried and eleven free SH groups could be detected and divided into three classes with different reactivities. In 1 mol of liver VRS one buried and nine free thiol groups were determined and these could be divided only into two classes. These observations suggest the existence of two VRS isoenzymes with different radiosensitivities: the more sensitive occurs mainly in brain and the less sensitive mainly in the liver of chick embryo.

18-day-old chick embryos (Leghorn) were irradiated in vivo with 1,2,3,4,5 or 7 Gy 60Co gamma-rays (dose rate = 0.9 Gy/min). Twenty-four hours after irradiation the activity of valyl-tRNA synthetase isolated from the brains was determined and compared with that of the non-irradiated control. The aminoacylation activity was found to decrease exponentially as a function of the dose (D37 = 6 Gy). Irradiation caused a more pronounced decrease in valyl-tRNA synthetase activity in 18-day-old than in 14-day-old chick embryos and induced in the valine-dependent ATP-PPi exchange reaction less change than in the valyl-tRNA FORMATion. The exposure of 15-day-old chick embryos to a dose of 4 or 5 Gy induced, respectively, a 40 or 20 per cent increase in enzyme activity of VRS (prepared on day 19 of embryonic life) relative to the control. In these experiments a decrease of Km value for tRNA has been found. The change in the number of sulfhydryl groups was also investigated.

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Investigations have been carried out concerning the effects of microwave (MW) exposure on the aminoacyl-transfer ribonucleic acid (tRNA) synthetase of the progeny of females that were exposed during their entire period of gestation (19 days). The changes caused by continuous-wave (CW) and amplitude-modulated (AM) MW radiation have been compared. CFLP mice were exposed to MW radiation for 100 min each day in an anechoic room. The MW frequency was 2.45 GHz, and the amplitude modulation had a 50 Hz rectangular waveform (on/off ratio, 50/50%). The average power density exposure was 3 mW/cm2, and the whole body specific absorption rate (SAR) was 4.23 +/- 0.63 W/kg. The weight and mortality of the progeny were followed until postnatal day 24. Aminoacyl-tRNA synthetase enzymes and tRNA from the brains and livers of the offspring (461 exposed, 487 control) were isolated. The aminoacyl-tRNA synthetase activities were determined. The postnatal increase of body weight and organ weight was not influenced by the prenatal MW radiation. The activity of enzyme isolated from the brain showed a significant decrease after CW MW exposure, but the changes were not significant after 50 Hz AM MW exposure. The activity of the enzyme isolated from liver increased under CW and 50 Hz modulated MW.