18S rRNA Synthesis Research Articles

Ribonucleic acid synthesis in embryonic chick muscle, rates of synthesis and half-lives of transfer and ribosomal RNA species

In this study the term 'rates of synthesis' does not refer to rates of transcription but to rates of accumulation of newly synthesized RNA. The rates of synthesis and half-lives of tRNA, 18S and 28S rRNA of 14-day and 17-day embryonic chick muscle were determined by following the accumulation of radioactive AMP in RNA after administration of [3H]8-adenosine to chick embryos and correcting for the average specific activity of the precursor ATP pool Transfer RNA of 14-day embryonic chick muscle is synthesized at a rate of 2.4 x 10(5) molecules per min per 2N DNA content and decays with a half-life of 50h. Transfer RNA of 17-day embryonic chick muscle is synthesized, and decays, at similar rates. Ribosomal RNA, 18S and 28S, are synthesized at a rate of 9.94 x 10(3) and 8.48 x 10(3) molecules per min per 2N DNA, respectively, in 14-day embryonic chick muscle. The rates of rRNA syntheis in 17-day embryonic muscle are also similar. In both 14-day and 17-day embryonic muscle, 18S and 28S rRNA each decays with a half-life of 65h. We conclude that the constant level of tRNA and rRNA in embryonic chick muscle from 14 to 17 days (Nwagwu & Nana, 1974) is maintained also by a constant rate of synthesis and turnover.

Deletion mutants of Xenopus laevis 5S ribosomal DNA

Deletion mutants have been derived from a plasmid-cloned repeating unit of Xenopus laevis oocyte 5S DNA by introducing the transposable chloramphenicol-resistance element Tn9 into the AT-rich spacer sequence near the 5′ terminus of the X. laevis 5S rRNA gene in a recombinant plasmid and then selecting plasmids which had lost the transposable element. Plasmids lacking the entire transposable element and various protions of the AT-rich spacer sequence flanking the original site of Tn9 integration have been obtained, and their ability to support transcription of the remaining X. laevis 5S rRNA gene has been tested in X. laevis oocyte nuclei. The deletion mutants analyzed in the present study retain the 49 nucleotide nonrepetitive sequence immediately adjacent to the 5′ terminus of the gene, but lack as much as 80% of the repetitive AT-rich spacer sequence ( Fedoroff and Brown, 1978). Such deletion mutants are fully active templates for 5S rRNA synthesis. This implies that the AT-rich spacer, which comprises half or more of each repeating unit in X. laevis oocyte 5S DNA, is relatively unimportant for correct initiation of transcription, and that if there are extragenic sequences with promoter function, they are likely to reside in the short nonrepetitive region immediately adjacent to the gene.

Zur synthese der plastidenribosomen während der leukoplasten-chloroplasten-transformation in callusgeweben von Nicotiana tabacum Synthesis of plastid ribosomes during leucoplast-chloroplast-transformation in tissue cultures of Nicotiana tabacum

Synthesis of plastid ribosomes was studied following synthesis of 5s rRNA of plastid ribosomes in tissue cultures of Nicotiana tabacum . It could be shown that 5s rRNA of plastid ribosomes is synthesized during the first hours after illumination. Synthesis of membranes, chlorophyll, and photosynthetic enzymes shows a lag-phase of one or two days after illumination. As no plastid ribosomes could be found in dark grown tissue cultures it is concluded that after illumination first plastid ribosomes have to be synthesized before leucoplast-chloroplast-transformation will start.

The regulation of RNA synthesis in yeast II: Amino acids shift-up experiments

A study has been made of the effects of a casamino acids shift-up on a prototrophic strain of yeast growing under conditions of ammonium repression. The shift-up produced an increase in growth rate some 120 min after the addition of amino acids to the medium. This growth rate increase was slightly preceded by an increase in the rate of accumulation of DNA. In contrast, the rate of accumulation of protein increased immediately and that of RNA 15-20 min after the shift. RNA was initially accumulated at a rate greater than that required to sustain the new steady state. This was shown to be due to an increase in the rate of synthesis of the rRNA species derived from the 35S precursor. The rate of synthesis of 5S rRNA and of tRNA increased much later and to a lesser extent than that of the 35S derived species. The implications of these results for general theories of regulation of RNA synthesis are discussed.

The metabolism of high-molecular-weight ribonucleic acid including polyadenylated species, in the developing rat brain

High-molecular-weight RNA was isolated from rat brain at various times after the intracranial administration of [32P]Pi. The synthesis of 28S and 18S rRNA could be detected within 1h of the injection of the radioactive precursor and appeared to be more pronounced, relative to other high-molecular-weight RNA, in the brains of older rats compared with those of newborn rats. Polyadenylated RNA, representing most mRNA and their precursors, was isolated by chromatography on oligo(dT)-cellulose. The contribution of this polyadenylated RNA to total RNA synthesis was investigated in the cerebral cortex and the phylogenetically older brain stem at different stages in the development of the rats by using a 5h period of labelling as an arbitrary index of transcription. In the brain stem the proportion of labelled polyadenylated RNA comprised 27-30% of the total RNA. The corresponding values for the cortex decreased from 34% in newborn rats to 23% in 40-150-day-old rats. These data indicated that proportionately more polyadenylated RNA is synthesized in the cortex of the newborn than in the adult rat and that there is a progressive decrease in the synthesis of polyadenylated RNA relative to rRNA during development.

Inhibitor of ribosomal RNA synthesis in Xenopus laevis embryos : V. Inability of inhibitor to repress 5S RNA synthesis

A specific inhibitor of ribosomal RNA (rRNA) synthesis was partially purified from an acid-soluble fraction of Xenopus laevis blastulae. Effects of this inhibitor on 5S rRNA synthesis of isolated neurula cells of the same species were investigated. The results show that the synthesis of both 5S rRNA and 4S RNA proceeds normally when both 18 and 28S rRNA are almost completely inhibited. Failure of the inhibitor to suppress 5S rRNA synthesis suggests that it plays an important role in the regulation of 18 and 28S rRNA synthesis during development and that the synthesis of 5S rRNA is not coordinated to that of 18 and 28S rRNA.

The role of 5-S RNA in temperature-sensitive ribosomal RNA maturation

The synthesis of 5-S RNA was found to be unchanged at both the permissive (33.5°C) and non-permissive (38.5°C) temperatures in a temperature-sensitive Baby Hamster Kidney cell line (BHK 21 ts 422E) as measured relative to synthesis of 18-S rRNA. The 5-S RNA is shown to be associated with nucleolar ribonucleoprotein particles even though rRNA processing does not yield a functional 28-S rRNA at the non-permissive temperature. The amount of 5-S RNA found associated with the 80-S ribonucleoprotein particles was the same at the permissive and non-permissive temperatures, indicating that an aberrant 5-S RNA contribution to rRNA processing is not a primary cause for the temperature-sensitive lesion of rRNA maturation in this mutant cell line. The amount of 5-S RNA in nucleolar 80-S RNA particles indicated that the association of 5-S RNA with the rRNA precursor particle occurs before the cleavage step at which 32-S precursor RNA is produced.

THE SYNTHESIS OF 5S RIBOSOMAL RNA DURING POLLEN DEVELOPMENT.

Tradescantia paludosa 5S ribosomal RNA (rRNA) has been characterized with respect to its base composition and relative electrophoretic mobility in comparison with that of E. coli. The period of 5S rRNA synthesis during pollen grain development was determined by pulse labeling the RNA synthesized during a 24 hr period of development with 32 P and then chasing in cold medium until pollen maturity. The period of highest 5S rRNA synthesis was found to occur prior to microspore mitosis. During and following mitosis over a period of 4 days there was a sharp decrease in the amount of 5S RNA synthesized and during the last 48 hr of pollen maturation, no 5S rRNA was synthesized.

Synthesis of ribonucleic acid in the venom gland of Crotalus durissus terrificus (Ophidia, Reptilia) after manual extraction of tne venom.

1. The incorporation of [5-(3)H]uridine into RNA of the venom gland of Crotalus durissus terrificus was studied after manual extraction (;milking') of the venom. The labelled precursor was injected immediately after milking. 2. The RNA was extracted 1, 2, 4, 6 and 8h after injection of the label and analysed by sucrose-density-gradient centrifugation. 3. The sedimentation analysis showed that 18S rRNA synthesis is higher than 28S rRNA at all time-intervals. The specific radioactivities of both ribosomal components did not reach a plateau even at 8h after injection. 4. An RNA fraction was detected sedimenting between 18S rRNA and 4S tRNA and was called the 10-14S fraction. The specific radioactivity was always higher than that of both classes of rRNA and reached the maximum value at 6h of labelling. 5. The incorporation of the precursor was also studied by radioautography, which helped to elucidate the intracellular origin of the RNA analysed by sucrose-density-gradient centrifugation.

Coordination of ribosomal RNA synthesis in vertebrate cells.

AbstractXenopus embryo cells and HeLa cells were investigated under various conditions to test for coordinate synthesis of high molecular weight (28S and 18S) and low molecular weight (5S) rRNA. Xenopus embryos initiate 28S and 18S rRNA synthesis at gastrulation (Brown and Littna, '64); we found that 5S rRNA synthesis is coordinately initiated with the 28S and 18S rRNAs at the same time in development. Dissociated Xenopus blastula cells were cultured in vitro for several hours to condition the medium; post‐gastrula cells were then grown in the conditioned medium to test for the existence of an inhibitor of rRNA synthesis. No inhibitor was detected.Low doses of actinomycin D profoundly inhibit the synthesis of 28S and 18S rRNA in HeLa cells, while 5S rRNA synthesis is less affected by this treatment. Therefore, actinomycin D does not produce a coordinate inhibition of all rRNA species. Similar effects of the antibiotic were found in cultured amphibian cells. Synchronized HeLa cells reinitiating RNA synthesis following mitosis also respond to actinomycin D in a non‐coordinate manner.