Wheat embryo ribonucleates. VIII. The presence of 7-methylguanosine 'cap structures' in the RNA of imbibing wheat embryos

1977 ◽  
Vol 55 (8) ◽  
pp. 819-824 ◽  
Author(s):  
M. S. Saini ◽  
B. G. Lane

1. By imbibing wheat embryos in media that contain methyl-labelled methionine, it is possible to label both terminal and nonterminal 7-methylguanosine constituents in NaCl-insoluble (2.5 M, 0 °C) RNA (iRNA).2. Most of the 7-[Me-14C]methylguanosine in wheat embryo i[Me-14C]RNA is present in nonterminal positions of polynucleotide chains, probably in ribosomal RNA.3. By passage through a column of oligo-dT-cellulose, it is possible to show that most of the 7-[Me-3H]methylguanosine in a 'bound' fraction of i[Me-3H]RNA from imbibing wheat embryos is present in terminal 'cap' structures, probably in messenger RNA.4. Although most of the 7-[Me-3H]methylguanosine in the 'unbound' (to oligo-dT-cellulose) fraction of i[Me-3H]RNA was present in nonterminal positions, there was also a highly significant fraction of 7-[Me-3H]methylguanosine in terminal 'cap' structures. Although it will be a subject of continued investigation, possible reasons why a large fraction of the total 7-[Me-3H]-methylguanosine was present in the 'unbound' fraction, in this present study, are a subject of discussion.5. Careful analysis failed to reveal the presence of any N6,O2′-di[Me-3H]methyladenosine in the 'unbound' fraction of i[Me-3H]RNA.6. Factors that might influence the binding of 'cap' oligonucleotides to DEAE-cellulose are the subject of a brief discussion.

1978 ◽  
Vol 56 (3) ◽  
pp. 197-202 ◽  
Author(s):  
T. D. Kennedy ◽  
B. G. Lane

There are conserved complements of ribosomal RNA (rRNA), transfer RNA (tRNA), and messenger RNA (mRNA) in dry wheat embryos. Although early labelling of RNA is largely directed toward the synthesis of complete molecules of nascent rRNA and mRNA, there is also selective labelling at 3′-hydroxyl termini in conserved polynucleotides when dry wheat embryos are subjected to short-term (0.5 h) imbibition in a medium that contains tritium-labelled adenosine, guanosine, cytidine, and uridine. Conserved tRNA is the principal mass component in NaCl-soluble RNA (sRNA) and most of the 'rapid labelling' of sRNA (rl-sRNA) is a result of labelling at 3′-hydroxyl termini in conserved tRNA. By contrast, although conserved rRNA is the principal mass component in NaCl-insoluble RNA (iRNA), most of the labelled 3′-hydroxyl termini in 'rapidly labelled' iRNA (rl-iRNA) do not appear to derive from rRNA. Although about 10% of the labelled 3′-hydroxyl termini in rl-iRNA originates in conserved poly(A)-rich mRNA, the available evidence leads to the conclusion that most of the labelled 3′-hydroxyl termini in rl-iRNA originates in an unusual NaCl-insoluble fraction of conserved tRNA. During the course of extended imbibition, between 0.5 and 5 h, there are characteristic changes in the chain lengths and labelling patterns for 3′-hydroxyl terminal poly(A) sequences in mRNA. Analytical and physiological implications of these data are subjects of discussion.


1981 ◽  
Vol 59 (10) ◽  
pp. 868-870 ◽  
Author(s):  
Byron G. Lane

Although template-active RNA in dry seeds and embryos has attracted widespread interest, there have been no published reports about 5′-terminai "capping" sequences in such RNA. Boro[3H]hydride labeling of periodate-oxidized termini and high performance liquid chromatography of cap oligonucleotides have been used to compare terminal sequences in poly(A)-rich RNA from dry and germinating embryos. As is the case in germinating embryos, poly(A)-rich RNA from dry embryos contains only "type 0" cap sequences, i.e., m7G(5′)ppp(5′)N, in which m7G is the 7-methylguanosine cap and N is any of the classical ribonucleosides: adenosine (A), guanosine (G), cytidine (C), and uridine (U). Striking differences between the cell-free translational capacities of bulk messenger RNA (mRNA) populations from dry and germinating embryos are not reflected in signal differences in their proportions of "type 0" cap structures: in general, there is approximately 40% m7G(5′)ppp(5′)A, with roughly equivalent amounts of m7G(5′)ppp(5′)G and m7G(5′)ppp(5′)C accounting for most of the remaining sequences. The findings with mRNA from dry plant embryos serve to emphasize interesting differences between patterns of methylation in the capped and uncapped RNA molecules in higher plants and animals; these differences have not been previously noted in the literature and are the subject of brief comment in this paper.


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)


1979 ◽  
Vol 57 (9) ◽  
pp. 1170-1175 ◽  
Author(s):  
A. C. Cuming ◽  
T. D. Kennedy ◽  
B. G. Lane

Commercially milled wheat germ is shown to be a convenient source material for facile recovery of mass (milligram) quantities of highly purified poly(A)-rich RNA. This poly(A)-rich RNA is efficiently translated in a nuclease-treated extract of rabbit reticulocytes. By sucrose density gradient fractionation of bulk poly(A)-rich RNA from wheat germ, it has been possible to show that there is a direct relationship between the molecular weights of the polypeptide products of cell-free synthesis and the molecular weights of the wheat mRNA molecules which program their synthesis. As assessed by SDS – polyacrylamide gel electrophoresis, the same array of polypeptides is synthesized when nuclease-treated reticulocyte extract is programmed by poly(A)-rich RNA from either commercially supplied or laboratory-prepared wheat embryos. Significantly, there are gross quantitative if not qualitative differences between the translational capacities of poly(A)-rich RNA from dry and imbibing wheat embryos, and the possible importance of these differences for interpreting a changing pattern of polypeptide synthesis in imbibing wheat embryos is the subject of a brief discussion.


1990 ◽  
Vol 124 ◽  
pp. 451-455
Author(s):  
L. Colina ◽  
W. Sparks ◽  
F. Macchetto

Despite intense theoretical and observational effort during the past few years, two important questions in the field of active galaxies remain open and are the subject of vigorous debate.The first concerns the connection between the interaction/merger of galaxies and the generation of starburst and/or active nuclei. A large fraction of high luminosity radio galaxies (Baum et al. 1988), quasars (Hutchinga, 1987) and IRAS galaxies (Sanders et al. 1988), show the presence of a large companion or have a peculiar optical morphology which is offen interpreted as evidence of a recent merger or interaction. In Seyferts, there are also several examples of individual galaxies with clear evidence of interaction/merger (Colina et al. 1987; Fricke and Kollatschny, 1989; Macchetto et al. 1990). Also a significant fraction of low luminosity radio galaxies are associated with a system of two elliptical galaxies in interaction (Colina and Pérez-Fournon, 1990a,b).


1973 ◽  
Vol 51 (5) ◽  
pp. 606-612 ◽  
Author(s):  
A. A. Azad ◽  
B. G. Lane

(1) When wheat embryos are extracted with aqueous phenol and the aqueous phase is made 2.5 M with respect to NaCl at 0 °C, there is selective precipitation of about 80% of the total RNA. The 18 S and 26 S RNA species from the wheat-embryo ribosomes comprise a preponderant mass fraction (ca. 80%) of this NaCl-insoluble RNA (iRNA). A small amount of a rapidly migrating electrophoretic component (iRMEC) can be released by aqueous denaturation of wheat-embryo NaCl-insoluble RNA and because it is specifically complexed with 26 S RNA, the iRMEC component has been termed a "satellite" of 26 S RNA.(2) The wheat-embryo satellite RNA has been shown to be present in the microsomal fraction recovered from cell-free homogenates of wheat embryos.(3) The wheat-embryo satellite RNA has been shown to be differentially localized in the large subunit of wheat-embryo ribosomes where it presumably exists as part of the same intermolecular 26 S RNA complex that can be isolated by directly extracting the whole embryos with aqueous phenol.(4) During preparation of the ribosomal subunits, there is substantial degradation of the component ribonucleates and the nature of this degradation is the subject of a brief 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.


2020 ◽  
Vol 65 (2) ◽  
pp. 297-319
Author(s):  
Aluaș Alina

"The Theatrical Potential in David Foenkinos’ Work. Analysis of the Novel, the Scenario and the Film “La Délicatesse”. Our interest, especially when it comes to the subject of literature, is to show the manner in which the text processing done by the author (script writer/director) brings to light the guidelines of the novelistic text’s semantics, which under careful analysis reveals a kind of personal myth of the novelist. The skewed, syncopated, interrupted writing which disrupts the chronotope serves the needs of the script as well as the director’s selective vision. Unconsciously, the novel seems to follow the structure of the theatrical model. These traits can also be found in the cinematographic structure of the film. Keywords: love, eroticism, delicacy, theatricality, scenario, film. "


1982 ◽  
Vol 60 (3) ◽  
pp. 389-397 ◽  
Author(s):  
Zbyszko F. Grzelczak ◽  
Mark H. Sattolo ◽  
Linda K. Hanley-Bowdoin ◽  
Theresa D. Kennedy ◽  
Byron G. Lane

The most prominent methionine-labeled protein made when cell-free systems are programmed with bulk mRNA from dry wheat embryos has been identified with what may be the most abundant protein in dry wheat embryos. The protein has been brought to purity and has a distinctive amino acid composition, Gly and Glx accounting for almost 40% of the total amino acids. Designated E because of its conspicuous association with early imbibition of dry wheat embryos, the protein and its mRNA are abundant during the "early" phase (0–1 h) of postimbibition development, and easily detected during "lag" phase (1–5 h), but they are almost totally degraded soon after entry into the "growth" phase of development, by about 10 h postimbibition.The most prominent methionine-labeled protein peculiar to the cell-free translational capacity of bulk mRNA from "growth" phase embryos is not detected as a product of in vivo synthesis. Its electrophoretic properties and its time course of emergence, after 5 h postimbibition development, suggest that this major product of cell-free synthesis may be an in vitro counterpart to a prominent methionine-labeled protein made only in vivo, by "growth" phase embryos. Designated G because of its conspicuous association with "growth" phase development, the cell-free product does not comigrate with any prominent dye-stained band in electrophoretic distributions of wheat proteins. The suspected cellular counterpart to G, also, does not comigrate with a prominent dye-stained wheat protein during electrophoresis, and although found in particulate as well as soluble fractions of wheat embryo homogenates it is not concentrated in either nuclei or mitochondria, as isolated.


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