Factors affecting protein synthesis in rat kidney ribosome preparations

Nicholls, D.M.; Ryan, M. P.; Miall, Stephen; Cappon, I.D.

doi:10.1139/o70-017

Cited by 14 publications

(11 citation statements)

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“…The validity of the methods on which this conclusion is based may be inferred from their suitability when used to study RNA synthesis in Xenopus embryos (20) and in mouse kidney (13), an organ rich in nucleases (31,32). The sensitivity of the methods permits detection of an estimated-200-600 molecules of 45S RNA turning over in 3-7 min per mouse kidney nucleus (13), but its sensitivity for the more slowly turning over RNA of toad bladder has not yet been estimated.…”

Section: Xf)mentioning

confidence: 99%

Kinetics of RNA synthesis in toad bladder epithelium: action of aldosterone during the latent period

Vancura¹,

Sharp²,

Malt³

1971

J. Clin. Invest.

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A B S T R A C T Processing of RNA in the toad bladder was analyzed by polyacrylamide-gel electrophoresis to determine whether aldosterone causes any changes in the 1 hr before it potentiates transport of sodium ion. No change was found in the quantity or in the specific activity of bulk RNA labeled with uridine-5-3H. In vivo and in vitro with either uridine-5-'H or with methionine-(methyl)-'H as precursors, processing of RNA was extremely slow. Heterodisperse RNA was obvious after 30 min of continuous labeling, but labeling of the 40S precursor of ribosomal RNA was not apparent for 60 min. Labeling of mature 28S and 18S RNA first became apparent after 8 hr. -S RNA was the principal fastmigrating species labeled at 30 min, and 4S RNA was not heavily labeled until 1 hr. Aldosterone (5 x 10-mole/liter) produced no changes. If care were not taken to inhibit metabolism of native bacteria colonizing the bladder, bacterial RNA of high specific activity predominated. We conclude that RNA metabolism in the toad bladder is extraordinarily slow, that a major acceleration of de novo synthesis in response to physiologic doses of aldosterone was not demonstrable, and that some reports to the contrary may have been influenced by artifacts from bacterial RNA metabolism. Earlier evidence for obligatory alterations in RNA metabolism during the latent period is not strong.

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Section: Xf)mentioning

confidence: 99%

Kinetics of RNA synthesis in toad bladder epithelium: action of aldosterone during the latent period

Vancura¹,

Sharp²,

Malt³

1971

J. Clin. Invest.

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“…The addition of 19 exogenous amino acids (exclusive of leucine), at a concentration of 5 pM, did not increase the incorporation of 14C-leucine, suggesting that the ribosomes and pH 5 fraction contained amino acids in adequate amounts as has been shown for other eucaryotic systems (12)(13)(14).…”

Section: Resultsmentioning

confidence: 56%

“…Radioactivity Protein was determined by the method of Lowry as described by NicholIs et al (12) with bovine serum albumin (Sigma) as a standard. All protein determinations were done in duplicate.…”

Section: Determination Of Protein and Measurement Ofmentioning

confidence: 99%

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Effects of puromycin aminonucleoside on protein synthesis in Absidia coerulea

Nicholls¹,

Cohen²

1970

Can. J. Biochem.

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NICHBLLS, D. M., AND COHEN, J. H. Effects of puromycin aminonucleoside on protein synthesis in Absidia coerukea. Can. J. Bioehem. 48,[858][859][860][861][862].The incorporation of '4C-leucine, '4C-phenylalanine, and '4C-phenylalanyB-taNA into protein was measured in-ribosome preparations of the soil fungus Absidia coeruka. The incorporation depended on ribosomes and GTP, together with pH 5 fraction and an ATP-generating system, or 1x11 sap. The addition of 5 x 1 8 -W aan%inonucleoside to the fungal cultures in vivo decreased the subsequent incorporation in vitro except when the ribosomes were prepared with bentonite. The percentage dry weight and protein concentration in the 140 080 x g supernatant were decreased (85 % and 30%, respectively) following addition of aminonucleoside in vivo. Preincubation of the cell-free system with 5 x los5 M or 5 x A4 aminonucleoside caused a decrease in the. subsequent incorporation of 14C-leucine and 14C-phenylalanyl-tRNA into protein. IrmtPoduetisnThe antibiotic puromycin inhibits protein synthesis in 70 S and also in 80 S ribosome systems (1). The inhibition results from puromycin acting as an analogue of aminoacyl-tRPJA2 so that the growing peptidyl group is transferred from peptidyl-tRNA to puromycin rather than to the amino group of the next aminoacyl-tRNA, thus ending the extension of the polypeptide chain (2). The puromycin reaction has been extensively studied and has provided much information on protein biosynthesis (3).The aminonucleoside of puromycin (6-N-added to cell-free preparations has not been found to have an effect on the incorporation of amino acids into protein in bacterial preparations (4). Several workers have reported that AN added to cultures of bacteria or mammalian cells caused an inhibition of WNA synthesis, particularly of ribosomal RNA (5-8). Since a large effect on protein synthesis was found in cell-free kidney and liver preparations obtained following the development of the characteristic ANinduced nephrotic syndrome in rats (9), it was desirable to see whether AN would have an effect on protein synthesis in cell-free preparations obtained from other eucaryotic cells such as those of fungi. lPresent address: Materials and Methods C~ilturesAn inoculum of the soil fungus Absidia coendlea (belonging to the Mucoraceae) was obtained by transferring mycelium that was growing on a plain agar plate culture. The plates were obtained by inoculating 1 % plain agar with a portion of the mycelium growing om potato dextrose agar (Difco) plate culture. The plate cultures were grown for a week at room temperature and then stored for 1-3 months at 2-4".Submerged cultures of the fungus were grown at 22-24" with shaking in 250-1111 flasks containing 125 ml medium of the following composition: 1.5 % glmcaase, 1 "/, inycological peptone (Oxoid), 0.05% magnesium sulfate, and 0.1 % K2HP04. Retardation of growth occurred at lower temperatures and ceased at 2-4". AN was added once to the cultures 24 h after inoculation at the beginning of the linear growth phase, and cul...

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“…leucine into proteins using PMS or microsomes of kidneys was performed by the method of Nicholls et al (5). Microsomes were prepared by centrifuging PMS for 1 hr at 40,000 rpm in a Spinco Model L3-40 ultracentrifuge.…”

Section: In Vitro [14c]leucine Incorporation Into Proteins In Vitro mentioning

confidence: 99%

Effect of Hypertonic Sodium Chloride on Polyribosomes and Protein Synthesis of Kidneys of Rats

Murty

Oliveros

Sidransky

1982

Experimental Biology and Medicine

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The effects of a single intraperitoneal administration of hypertonic NaCl(5.3 ml of 7.55% NaCYlOO g body wt) on polyribosomes and in vitro protein synthesis of kidneys of rats were investigated. The results revealed that there was marked disaggregation of kidney polyribosomes of rat (fasted or nonfasted) that received a single dose of hypertonic NaCl solution 30 min prior to sacrifice in comparison to control rats that received isotonic NaCl solution. In vitro [ 14C]leucine incorporation into protein was significantly decreased using either postmitochondrial supernatant or microsomes from kidneys of hypertonic NaCltreated rats compared with control rats. Administration of a single dose of hypertonic NaCl to rats produced a significant increase in the activity of renal RNase in comparison to that of the control group. Assay of renal acid phosphatase in the particulate and soluble fractions revealed no changes in the activities in the control and experimental rats. 258

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Factors affecting protein synthesis in rat kidney ribosome preparations

Cited by 14 publications

References 1 publication

Kinetics of RNA synthesis in toad bladder epithelium: action of aldosterone during the latent period

Kinetics of RNA synthesis in toad bladder epithelium: action of aldosterone during the latent period

Effects of puromycin aminonucleoside on protein synthesis in Absidia coerulea

Effect of Hypertonic Sodium Chloride on Polyribosomes and Protein Synthesis of Kidneys of Rats

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