Effects of ribosomal wash factors and spermidine on endogenous and exogenous mRNA stimulated protein synthesis in the wheat germ cell-free system

1979 ◽  
Vol 57 (5) ◽  
pp. 429-435
Author(s):  
Elahe Elahi ◽  
George H. Jones
Keyword(s):  
Protein Synthesis ◽  
Germ Cell ◽  
Wheat Germ ◽  
Mrna Translation ◽  
The Other ◽  
Globin Mrna ◽  
Free System ◽  
Differential Effects ◽  
Myeloma Proteins ◽  
Dependent Protein

Differential effects of Mg2+, spermidine, and reticulocyte ribosomal wash factors on the translation of endogenous, myeloma, and globin mRNA's have been observed in studies with the wheat germ cell-free protein synthesizing system. Spermidine stimulated globin mRNA translation but not the translation of endogenous wheat germ messages, and the polyamine actually inhibited the translation of myeloma mRNA. Ribosomal wash factors, on the other hand, stimulated endogenous and myeloma mRNA dependent protein synthesis in an Mg2+-dependent fashion but inhibited globin mRNA translation. The combination of ribosomal wash factors and spermidine was either stimulatory or inhibitory depending on the Mg2+ concentration and the message. It was further observed that translation of exogenous myeloma mRNA proceeded for only 60 min at 25 °C under all conditions tested in this study, while translation of endogenous wheat germ messages continued for longer periods of time. No differential effects of spermidine on the synthesis of high molecular weight myeloma proteins were observed.

Rat b/b Anemia: Translation of Normal and Anemic Globin mRNA in Wheat-Germ Cell-Free System

1976 ◽  
Vol 71 (1) ◽  
pp. 85-91 ◽  
Author(s):  
Radomir CRKVENJAKOV ◽  
Slavko CUSIC ◽  
Ivan IVANOVIC ◽  
Vladimir GLISIN
Keyword(s):  
Germ Cell ◽  
Wheat Germ ◽  
Globin Mrna ◽  
Free System ◽  
Cell Free System

scholarly journals Evidence That Phosphorylation of the α-Subunit of eIF2 Does Not Essentially Inhibit mRNA Translation in Wheat Germ Cell-Free System

2020 ◽  
Vol 11 ◽  
Author(s):  
Andrey V. Zhigailov ◽  
Alena M. Alexandrova ◽  
Anna S. Nizkorodova ◽  
Gulshan E. Stanbekova ◽  
Ruslan V. Kryldakov ◽  
...  
Keyword(s):  
Germ Cell ◽  
Wheat Germ ◽  
Mrna Translation ◽  
Α Subunit ◽  
Free System ◽  
Cell Free System

Methylglyoxal inhibits the translation of natural and chemically decapped mRNAs

1984 ◽  
Vol 4 (9) ◽  
pp. 783-788 ◽  
Author(s):  
Andres M. Lozano ◽  
Vasek A. Mezl
Keyword(s):  
Protein Synthesis ◽  
Germ Cell ◽  
Chemical Reaction ◽  
Wheat Germ ◽  
Free System ◽  
Weak Inhibitor ◽  
Cell Free System ◽  
General Inhibitor ◽  
Dependent Inhibition ◽  
Reticulocyte Lysate

Methylglyoxal was a weak inhibitor of translation in the reticulocyte-lysate cell-free system and it did not display cap-dependent inhibition. A similar inhibition was obtained in a wheat-germ cell-free system that displayed extensive cap-dependent inhibition with the cap analogue 7-methylguanosine phosphate. These results show that the chemical reaction of methyl-glyoxat with 7-methylguanosine is not the mechanism for the inhibition of protein synthesis by methylglyoxal and that methylglyoxat is a weak general inhibitor of translation.

scholarly journals Inhibition of cell-free protein synthesis by low-molecular-weight nuclear polyadenylate-containing ribonucleic acid species isolated from the lactating guinea pig

1980 ◽  
Vol 186 (2) ◽  
pp. 561-570 ◽  
Author(s):  
I C Bathurst ◽  
R K Craig ◽  
P N Campbell
Keyword(s):  
Protein Synthesis ◽  
Mammary Gland ◽  
Germ Cell ◽  
Inhibitory Activity ◽  
Wheat Germ ◽  
Free System ◽  
Cell Free System ◽  
Mechanism Of Inhibition ◽  
Endogenous Activity ◽  
Cell Free Protein Synthesis

1. Poly(A)-containing RNA was isolated from the nuclei of mammary gland, liver and brain of lactating guinea pigs. 2. Total nuclear poly(A)-containing RNA from mammary gland inhibited mRNA-directed protein synthesis by a wheat-germ cell-free system. It also inhibited the endogenous activity of the wheat-germ and other cell-free systems. It did not inhibit a wheat-germ cell-free system directed by poly(U). 3. Total nuclear poly(A)-containing RNA from liver and brain did not inhibit the mRNA-directed wheat-germ system. 4. Fractionation of the nuclear poly(A)-containing RNA revealed inhibitory activity in the less than 10 S fraction from mammary gland as well as that from liver and brain. 5. The mechanism of protein-synthesis inhibition appeared to be at the level of elongation. 6. The inhibitory activity could be reversed in a wheat-germ system by increasing the amount of S-30 supernatant. 7. The mechanism of inhibition of protein synthesis is discussed in relation to other RNA species known to inhibit such systems.

Initiation in einem Polyribosomen-abhängigen Protein-synthetisierenden zellfreien System aus Saccharomyces / Initiation in a Polyribosome-Dependent Protein-Synthesizing Cell-Free System from Saccharomyces

1976 ◽  
Vol 31 (3-4) ◽  
pp. 169-173 ◽  
Author(s):  
Bernd Schulz-Harder ◽  
Ernst-Randolf Lochmann
Keyword(s):  
Protein Synthesis ◽  
Free System ◽  
Cell Free System ◽  
Aurintricarboxylic Acid ◽  
Dependent Protein ◽  
A Cell

Abstract A method to prepare polyribosomes from yeasts by using the french-press is described. The highest yield of polyribosomes was derived from late log-phase cells. These polyribosomes, incubated in a cell-free system, were able to reinitiate protein synthesis, which was shown by inhibiting aminoacid incorporation by aurintricarboxylic acid, edeine and sodiumfluoride. We developed the translational system in order to look for the optimal ion-conditions of a DNA-dependent protein-synthesizing system. We found out that at the optimal MgCL2-concentration (6 mᴍ) protein synthesis was strongly inhibited by Mangan ions which are required for transcription in yeast. If protein-synthesis was carried out with 2 mᴍ and 3 mᴍ MgCl2 maximal aminoacid incorporation was observed at 2 mᴍ and 1.5 mᴍ MnCl2.

scholarly journals Mechanism of action of purpuromycin

1992 ◽  
Vol 284 (1) ◽  
pp. 47-52 ◽  
Author(s):  
P Landini ◽  
E Corti ◽  
B P Goldstein ◽  
M Denaro
Keyword(s):  
Protein Synthesis ◽  
Rna Synthesis ◽  
Intact Cell ◽  
Acid Synthesis ◽  
Free System ◽  
Dna And Rna ◽  
Dna And Rna Synthesis ◽  
Dependent Protein ◽  
Elongation Step

Purpuromycin, an antibiotic active against both fungi and bacteria, shows different modes of action against these two kinds of micro-organisms; in Candida albicans it inhibits RNA synthesis, whereas in Bacillus subtilis protein synthesis is primarily affected, with DNA and RNA synthesis blocked at higher concentrations of the drug. In bacterial cell-free protein-synthesis systems, purpuromycin did not inhibit synthesis from endogenous mRNA (elongation of peptides initiated within the intact cell) but inhibited MS2-phase RNA-dependent protein synthesis (which requires initiation) by 50% at 0.1 mg/l. Poly(U)-directed polyphenylalanine synthesis was 50% inhibited by 20 mg of purpuromycin/l when added to a complete system; however, when purpuromycin was preincubated with ribosomes dissociated into 30 S and 50 S subunits, the concentration for 50% inhibition fell to 0.1 mg/l. By contrast, in a C. albicans cell-free system poly(U)-directed polyphenylalanine synthesis was partially inhibited only at 200 mg/l. Purpuromycin also inhibited polynucleotide synthesis in vitro in reactions using Escherichia coli or wheat-germ RNA polymerases or E. coli DNA polymerase I. We suggest that in bacteria the primary target of purpuromycin is on ribosomes and that its action precedes the elongation step of protein synthesis. The effect on nucleic acid synthesis in both fungi and bacteria may be due to interaction of purpuromycin with DNA.

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