Localization of the ribosomal RNA genes in Drosophila simulans

In situ hybridization of cloned rRNA genes from Drosophila melanogaster to D. simulans metaphase chromosomes shows that in the tested wild type strains both sex chromosomes contain a nucleolus organizer region. Silver grain counts support the published data that the X chromosomal rRNA gene number is significantly higher than the Y chromosomal.     

Organization of ribosomal RNA genes in Mycoplasma capricolum

Summary DNA segments carrying rRNA genes of Mycoplasma capricolum have been cloned and characterized by restriction endonuclease mapping, DNA-RNA hybridization and nucleotide sequencing. The M. capricolum genome has two sets of rRNA gene clusters, where the arrangement is in the order of (5)16S-23S-5S(3). The spacer region between 16S and 23S rDNA is extremely rich in AT and does not carry any tRNA genes. Present address: Division of Hematology and Immunology of Internal Medicine, Kanazawa Medical University, Uchinada-Cho, Kahoku-Gun Ishikawa Pref. 920-02, Japan     

Arrangement of the ribosomal RNA genes in chloroplast DNA of Leguminosae

Summary The circular chloroplast DNA from three species of plants in the taxonomic family Leguminosae were examined using electron microscopic techniques and restriction endonuclease digestion. Chloroplast DNAs from chickpea (Cicer arietinum), mung bean (Vigna radiata), and soy bean (Glycine max) were found to range in size from 119–151 kilobase pairs by contour length measurements. Sizes of the chloroplast DNAs have been further confirmed using different restriction endonucleases. Two of the chloroplast DNAs examined, soy bean and mung bean, contain a region approximately 15.9–18% of their monomer length that is repeated in reverse polarity. This repeated region separates a small unique region that ranges in size from 18.75–20.4 kilobase pairs and a large unique region that ranges in size from 73.4–85 kbp. This feature was not found in the chloroplast DNA of chickpea. R-loop hybridizations performed using chloroplast ribosomal RNAs demonstrate that the two ribosomal gene sets of the mung been and soy bean are arranged in inverted orientation within this repeated region. In contrast, the chickpea chloroplast DNA posesses a single ribosomal RNA gene set in the circular molecule. In all three chloroplast DNAs examined, the genes encoding the chloroplast 23S and 16S ribosomal RNA genes are separated by a spacer region which ranges in size from 2.2 to 2.48 kbp.     

Localization of ribosomal RNA genes by high resolution autoradiography

In the myxomycete Physarum polycephalum, the genes for ribosomal RNA replicate to a large extent in the G2 phase of the mitotic cycle. In mitotically synchronous plasmodia this fact allows the rRNA genes to be localized by electron microscopic autoradiography. After labelling with thymidine in the G2 phase, silver grains were concentrated over the fibrillar and not over the granular regions of the nucleoli. This shows the presence of DNA in these fibrillar regions or their immediate vicinity. Following labelling and chase during the G2 period, nucleoli are dispersed during mitosis and silver grains concentrate over some regions of condensed chromatin.     

Localization of ribosomal RNA genes on human acrocentric chromosomes

Summary Clonal derivatives of a human heteroploid cell line, with different numbers of acrocentric chromosomes, show different rDNA contents. A linear relationship has been found between the rDNA content and the relative mass of the acrocentric chromosomes (D+G) expressed as the ratio between the mass of their DNA and the mass of the DNA of the whole chromosomal complement. The results suggest that human rRNA genes are located exclusively on the chromosomes of the groups D and G and that all these chromosomes contain rRNA genes.     

Organization of the chloroplast ribosomal RNA genes of Euglena gracilis bacillaris

Summary The order of eight of the 29 endonuclease EcoRI-generated fragments of chloroplast DNA was determined. Three sets of rRNA genes aligned sequentially in the same orientation form part of this region. The repeated sets differ in the length and sequence of the spacers among themselves and with the rRNA genes of E. gracilis strain Z.     

Molecular cloning of the ribosomal RNA genes of the photosynthetic bacterium Rhodopseudomonas capsulata

Summary Chromosomal segments of Rhodopseudomonas capsulata carrying the ribosomal operons and cloned with the cosmid vector pHC79 have been identified by cross hybridization with ³²P-ATP labeled rRNAs. At least seven rRNA operons are present in the R. capsulata chromosome. By R-loop analyses of DNA-RNA hybrids, two distinct loop structures of sizes 1.50 kb and 2.52 kb corresponding to the 16S and 23S RNA molecules, respectively, were detected. Intact 23S RNA molecules can be isolated from R. capsulata ribosomes by sucrose density centrifugation. However, fragmentation of the 23S RNA molecule into a 16S-like molecule was observed during gel electrophoresis. Restriction mapping and hybridization of a 9 kb PstI fragment that contained one copy of the rRNA operon showed the following sequence of the RNA genes in R. capsulata 16S, 23S, and 5S. A spacer region of 0.91 kb was found between the 16S and the 23S RNA genes.     

Characterisation of the genes for ribosomal RNA in flax

DNA coding for the 18S and 25S rRNAs in flax (Linum usitatissimum) has been purified and is arranged in tandem arrays with a repeat length of 8.6 kb. There is no detectable variation in the size of this repeat unit. Single repeat units have been cloned in the plasmid pAT 153. The coding sequences for the 25S, 18s and 5.85 rRNAs have been localised by hybridisation. The cloned rDNA has been used to compare two genotrophs, L1 ad S1, where the number of rRNA cistrons has been altered by growth under different environmental conditions. In terms of the size of the repeat unit and the position of a number of restriction enzyme sites the rDNAs from L1 and S1 were identical.     

Control of ribosomal RNA synthesis in Escherichia coli

Summary The ribosomal RNA synthesis in a cell-free system containing the nucleoids and the cytoplasmic fraction prepared from Escherichia coli cells has been investigated. The addition of the 4S fraction from the cytoplasm to the isolated nucleoids induces RNA synthesis by a new chain initiation. In this system a preferential initiation of rRNA chains occurs. The experimental results suggest that the 4S fraction contains at least two activities, one for releasing RNA-polymerases from the nucleoids, and another for the frequent initiation of rRNA chains. No restriction of the rRNA synthesis has been observed in the nucleoids and the 4S fraction from the amino acidstarved rel ⁺ cells. The rRNA synthesized in the above system is detected at about 23S and 16S rRNA regions.     

Localization of the structural gene for ribosomal protein L19 (rplS) in Escherichia coli

Summary A temperature-sensitive mutant derived from an E. coli K12 strain, PA3092, was found to have an alteration in the ribosomal protein L19 (Isono et al., 1977). This mutant is a double mutant with a temperature-sensitivity mutation and a mutation leading to the structural alteration of L19 protein. Crosses with various Hfr strains and transductions with P1kc have revealed that the latter mutation maps at 56.4 min, between pheA and alaS. From the fact that two other mutations causing different types of alterations in L19 protein also map at this locus, the gene affected by these mutations was concluded to be the structural gene for the ribosomal protein L19 (rplS).     

Linkage of ribosomal RNA genes in Leptospira

We determined the linkage of 16S, 23S, and 5S rRNA genes in several strains of Leptospira and Leptonema by DNA-DNA hybridization. Almost all the hybridizations in all leptospires used in these experiments gave two radioactive bands and the results strongly suggest that the number of the 16S and the 23S rRNA genes in those strains is two, respectively. In contrast with the larger rRNAs, the number of 5S rRNA gene was different. In the strains of leptospires, L. biflexa, which were non-parasitic, there are two genes for 5S rRNA, whereas only one gene for 5S rRNA is carried in L. interrogans, which were originally isolated as parasitic. Southern hybridization experiments suggest that those rRNA genes are interspersed on the leptospiral chromosome.     

Saccharomyces cerevisiae mutants defective in the maturation of ribosomal RNA

Summary Slow-growing mutants were isolated after mutagenesis of the osmotic-sensitive strain Saccharomyces cerevisiae VY1160. The isolated mutants in rich media have generation times from 300 to 400 min at 30°C. Studies on the biosynthesis of rRNA^x have shown, that the processing of 37S pre-rRNA in 6 of the slow-growing mutants occurs 3 to 4 times slower than in the parental strain. These mutants with decreased rate of rRNA maturation are of two different types. In some of them the processing of both 37S and 27S pre-rRNA is slowed down, while the mutants from the second group are acharacterized by a specific inhibition of the step 27S pre-rRNA25S rRNA. Experiments in which the synthesis of macromolecules was studied, have shown that in the mutants and in the parental strain, RNA and proteins are synthesized at comparable rates. Preliminary results suggest that the decreased rate of rRNA processing in three of the isolated mutants might be due to an insufficient function of the enzymes involved in the maturation of rRNA.Abbreviations rRNA ribosomal RNA - pre-rRNA precursor to ribosomal RNA