Wheat embryo ribonucleates. VII. Rapid, efficient and selective formation of 5S–18S and 5.8S–26S hybrids in an aqueous solution of the four ribosomal polynucleotides, and the results of a search for the corresponding hybrids in wheat embryo ribosomes

1977 ◽  
Vol 55 (1) ◽  
pp. 99-109 ◽  
Author(s):  
K. M. Oakden ◽  
A. A. Azad ◽  
B. G. Lane

(1) If wheat embryo 5S and 5.8S rRNA are differentially labelled, it can be shown that there is highly selective association of 5S [14C]RNA with 18S rRNA, and of 5.8S [3H]RNA with 26S rRNA when a solution (0.3 M NaCl) that contains approximately equimolar amounts of the four ribosomal polynucleotides is heated briefly (3 min) at 60 °C.(2) Comparison of Tm values and melting profiles for laboratory-prepared and natural 5.8S–26S rRNA hybrids suggests that restoration of the natural union between 5.8S and 26S rRNA can be achieved with facility and fidelity in the laboratory.(3) Union between 5.8S and 26S rRNA remains intact when wheat embryo ribosomes are disintegrated either by digestion with pronase or by treatment with sodium dodecyl sulphate, but the same treatments release 5S and 18S rRNA as freely migrating electrophoretic components.(4) Intact 18S and 26S rRNA can be prepared from small and large subunits, respectively, when wheat embryo ribosomes are dissociated by treatment with 0.5 M KCl.(5) Incidental to the principal investigation, it has been shown that, even after storage for more than 6 years at − 70 °C, commercial supplies of roller-milled wheat germ yield S23 extracts that are very active in the cell-free translation of globin mRNA.(6) The physicochemical and possible biochemical significance of various types of intermolecular complexing between pairs of ribosomal polynucleotides is a subject of discussion.

1975 ◽  
Vol 53 (3) ◽  
pp. 320-327 ◽  
Author(s):  
A. A. Azad ◽  
B. G. Lane

Under the conditions used in this study, wheat-embryo 5S rRNA complexes with its homologous 18S rRNA from wheat embryos and with heterologous 18S rRNA from other eukaryotic source materials such as yeast, L cells, and HeLa cells, but it does not complex with heterologous 16S rRNA from a prokaryote such as Escherichia coli or with homologous or heterologous 26S(23S) rRNA of either eukaryotic or prokaryotic origin.If a solution of wheat-embryo rRNA is simply made 0.3 M with respect to NaCl and then heated at 60 °C for 3 min before quick cooling to room temperature (ca. 20 °C), there is both preferential and efficient complex formation between 5S and 18S rRNA and between 5.8S and 26S rRNA.The laboratory-prepared' complex between wheat-embryo 5S rRNA and its homologous 18S rRNA is more thermostable in 0.1 M NaCl solution than is the 'natural' complex between wheat-embryo 5.8S rRNA and its homologous 26S rRNA, and both complexes 'melt' over a narrow range of temperature.The possible physicochemical and physiological importance of both homologous and heterologous rRNA complexes is the subject of a brief discussion.


1977 ◽  
Vol 55 (5) ◽  
pp. 582-586 ◽  
Author(s):  
M. W. Gray ◽  
R. S. Cunningham

Two hypermodified, alkali-stable dinucleotide sequences, each containing abase modification in addition to sugar methylation, are known to be present in wheat embryo 26S + 18S rRNA (Gray, M. W. (1974) Biochemistry 13, 5453–5463). Quantitative analysis of unfractionated 26S + 18S rRNA had suggested that each of these sequences (Cm-ψp and ψm-Ap, where Cm = O2′-methylcytidine and ψm = O2′-methylpseudouridine) was present in either the 18S or the 26S rRNA species, but not in both, at a frequency of not more than once per chain. In the study reported here, the individual 32P-labeled 18S and 26S rRNA species were isolated from viable wheat embryos germinated in the presence of [32P]orthophosphate. From analyses of phosphodiesterase and alkaline hydrolysates of the separated [32P]RNAs, we conclude that ψm-Ap is confined to wheat cytosol 18S rRNA, whereas Cm-ψp is localized in wheat cytosol 26S rRNA. The presence of ψm in the 18S rRNA of wheat stands in contrast with the situation in animal cells, where this hypermodified nucleoside is located in the 28S rRNA (Khan, M. S. N. &Maden, B. E. H. (1976) J. Mol. Biol. 101, 235–254)


1982 ◽  
Vol 58 (1) ◽  
pp. 23-33
Author(s):  
R.L. Shoeman ◽  
H.G. Schweiger

Modification of existing two-dimensional techniques enables isoelectric focusing and sodium dodecyl sulphate polyacrylamide gel electrophoresis of complex mixtures of proteins to be completed within 8 h. The method was optimized to separate the protein components of a wheat germ cell-free translation system, providing a statistically proven resolution better than 0. 03 of a pH unit for the isoelectric point and 1000 for Mr. Fourteen of the more than 300 proteins separated were characterized with respect to Mr and isoelectric point relative to standard proteins under the same conditions. Stained wheat germ proteins thus serve as internal standards for analysis of in vitro translation products.


1978 ◽  
Vol 56 (6) ◽  
pp. 365-369 ◽  
Author(s):  
A. C. Cuming ◽  
B. G. Lane

It has been found that bulk poiy(A)-rich RNA from dry wheat embryos is broadly hetero-disperse when examined by polyacrylamide gel electrophoresis. The poly(A)-rich RNA from dry wheat embryos has been translated in a cell-free protein-synthesizing system from the same commerically supplied, roller-milled wheat embryos. Compatible with the electrophoretic heterodispersity observed for poly(A)-rich RNA, the radioactive products of its cell-free translation, when examined by sodium dodecyl sulphate polyacrylamide gel electrophoresis, have mobilities that are broadly coincident with the many dye-stained (nonradioactive) proteins present in wheat extracts. With due allowance for the limitations of the cell-free system, which is known to translate, selectively, lower molecular-weight species of mRNA, it has been concluded that the conserved poly(A)-rich mRNA in dry wheat embryos probably has the translational capacity required to account for the highly eclectic protein synthesis that we have observed during early (40-min) imbibition of viable wheat embryos.


1976 ◽  
Vol 54 (3) ◽  
pp. 261-271 ◽  
Author(s):  
K. M. Oakden ◽  
B. G. Lane

The NaCl-insoluble (2.5 M, 0 °C) fraction of wheat embryo RNA (iRNA) can be labelled when wheat embryos are subjected to either short-term (0.5 h) or long-term (24 h) imbibition in a medium that contains tritium-labelled adenosine, guanosine, cytidine and uridine. Electrophoretic analyses reveal that, after short-term labelling, there is a broadly heterodisperse distribution of radioactivity in 'rapidly labelled' i[3H]RNA, but after long-term labelling, there is an essentially trimodal distribution of radioactivity in i[3H]RNA. End-group analyses reveal that, after short-term labelling, adenosine is the principal 3′-hydroxyl terminus in all centrifugal subfractions of 'rapidly labelled' i[3H]RNA, whereas cytidine (in 5.8S rRNA), guanosine (in 18S rRNA) and uridine (in 26S rRNA) are the principal 3′-hydroxyl termini in centrifugal subfractions of wheat embryo i[3H]RNA. Guanosine is also the principal 3′-hydroxyl terminus in the 18S rRNA of differentiating embryos excized from both monocotyledonous (wheat, barley, corn) and dicotyledonous (pea) seedlings. The implications that the end-group measurements may have for current views about the possible biochemical involvements of 3′-hydroxyl terminal sequences in both mRNA and 18S rRNA are subjects of discussion. Incidental to the principal investigation, an existing technique for analyzing the RNA contents of cellular materials has been appropriately modified to circumvent interference from uv-absorbing pigments, which, when present, prevent application of the method to plant materials.


2002 ◽  
Vol 5 (6) ◽  
pp. 473-480
Author(s):  
Bentham Science Publisher A.N. Alexandrov ◽  
Bentham Science Publisher V.Yu. Alakhov ◽  
Bentham Science Publisher A.I. Miroshnikov

1978 ◽  
Vol 43 (4) ◽  
pp. 1184-1189
Author(s):  
Ota Fuchs ◽  
Jitka Borová ◽  
Přemysl Poňka ◽  
Jan Neuwirt

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yue Zhao ◽  
Qingyang Sun ◽  
Shusheng Zhu ◽  
Fei Du ◽  
Ruzhi Mao ◽  
...  

AbstractShangri-La is a wine region that has the highest altitude vineyards in China. This is the first study investigated the biodiversity of non-Saccharomyces yeasts associated with spontaneous fermentation of Cabernet Sauvignon wines produced from two sub-regions (Lancang River and Jinsha River) of Shangri-La. The culturable yeasts were preliminarily classified based on their colonial morphology on the Wallerstein Laboratory nutrient agar plates. Yeast species were identified by the sequencing of the 26S rRNA D1/D2 region and the 5.8S rRNA ITS region. Twenty-five non-Saccharomyces yeast species belonging to sixteen genera were isolated and identified in Shangri-La wine region. Candida, Hanseniaspora, Pichia, and Starmerella were found in both sub-regions, but the Lancang River showed more diverse yeast species than the Jinsha River. Shangri-La not only exhibited high diversity of non-Saccharomyces yeasts, and furthermore, seven species of non-Saccharomyces yeasts were exclusively found in this region, including B. bruxellensis, D. hansenii, M. guilliermondii, S. vini, S. diversa, T. delbrueckii and W. anomalus, which might play an important role in distinctive regional wine characteristics. This study provide a relatively comprehensive analysis of indigenous non-Saccharomyces yeasts associated with Cabernet Sauvignon from Shangri-La, and has significance for exploring ‘microbial terroir’ of wine regions in China.


Langmuir ◽  
2003 ◽  
Vol 19 (22) ◽  
pp. 9155-9161 ◽  
Author(s):  
Daniel Angelescu ◽  
Ali Khan ◽  
Horia Caldararu

1978 ◽  
Vol 79 (1) ◽  
pp. 132-137 ◽  
Author(s):  
G Mintz ◽  
L Glaser

After separation of whole proteins of chick neural retina by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS), a number of glycoproteins can be detected by staining the gels with 125I-labeled wheat germ agglutinin (WGA) and other lectins. The glycoprotein patterns show both quantitative and qualitative changes between days 7 and 13 of development. Some of these glycoproteins can be separated by chromatography on columns of insolubilized lectins. These observations suggest that purification of some of these glycoproteins identified by staining with radioactive lectins would yield retinal antigens which may be specific for developmental stage and cell type.


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