scholarly journals Unusual ciliate-specific codons in Tetrahymena mRNAs are translated correctly in a rabbit reticulocyte lysate supplemented with a subcellular fraction from Tetrahymena

1987 ◽  
Vol 244 (2) ◽  
pp. 331-335 ◽  
Author(s):  
P H Andreasen ◽  
H Dreisig ◽  
K Kristiansen
Keyword(s):  
Codon Usage ◽  
Genetic Code ◽  
Subcellular Fraction ◽  
Tetrahymena Thermophila ◽  
Rabbit Reticulocyte Lysate ◽  
Reading Frame ◽  
Universal Genetic Code ◽  
Reticulocyte Lysate ◽  
Translational Termination

The codon usage of Tetrahymena thermophila and other ciliates deviates from the ‘universal genetic code’ in that UAA and probably UAG are not translational termination signals but code for glutamine. Therefore, translation in vitro of mRNA from Tetrahymena in a reticulocyte lysate is prematurely terminated if a UAA or UAG triplet is present in the reading frame of the mRNA. We show that the addition of a subcellular fraction from Tetrahymena thermophila enables a rabbit reticulocyte lysate to translate Tetrahymena mRNAs into full-sized proteins. The activity of the subcellular fraction is shown to depend on the combined function of a protein component(s) and a tRNA(s). The subcellular fraction is easily prepared and its usefulness for the identification of isolated mRNAs from Tetrahymena by their translation products in vitro is demonstrated.

scholarly journals Seed extracts inhibiting protein synthesis in vitro

1980 ◽  
Vol 186 (2) ◽  
pp. 439-441 ◽  
Author(s):  
A Gasperi-Campani ◽  
L Barbieri ◽  
P Morelli ◽  
F Stirpe
Keyword(s):  
Protein Synthesis ◽  
Rabbit Reticulocyte Lysate ◽  
Intact Cells ◽  
Euonymus Europaeus ◽  
Lower Extent ◽  
Reticulocyte Lysate ◽  
Dialysis Membranes ◽  
Inhibiting Protein ◽  
Seed Extracts

Of 33 seed extracts examined, 12 inhibited protein synthesis in a rabbit reticulocyte lysate. This activity seems to be due to a protein, since (i) it was recovered with the (NH4)2SO4 precipitate, (ii) it was retained by dialysis membranes, and (iii) in all cases but one was destroyed by boiling. Only the extracts from the seeds of Adenia digitata and, to a lower extent, of Euonymus europaeus inhibited protein synthesis in intact cells.

In Vitro Cleavage of eIF4GI but not eIF4GII by HIV-1 Protease and its Effects on Translation in the Rabbit Reticulocyte Lysate System

2002 ◽  
Vol 318 (1) ◽  
pp. 9-20 ◽  
Author(s):  
Théophile Ohlmann ◽  
Déborah Prévôt ◽  
Didier Décimo ◽  
Florence Roux ◽  
Jérôme Garin ◽  
...  
Keyword(s):  
Rabbit Reticulocyte Lysate ◽  
Rabbit Reticulocyte Lysate System ◽  
Reticulocyte Lysate ◽  
Hiv 1

scholarly journals Assembly of the ribonucleoprotein complex containing the mRNA of the β-subunit of the mitochondrial H+-ATP synthase requires the participation of two distal cis-acting elements and a complex set of cellular trans-acting proteins

2002 ◽  
Vol 365 (2) ◽  
pp. 417-428 ◽  
Author(s):  
Javier RICART ◽  
José M. IZQUIERDO ◽  
Carlo M. Di LIEGRO ◽  
José M. CUEZVA
Keyword(s):  
Atp Synthase ◽  
Sucrose Concentration ◽  
Β Subunit ◽  
Reading Frame ◽  
Ribonucleoprotein Complex ◽  
Cis Acting ◽  
Translational Machinery ◽  
F1 Atpase ◽  
Reticulocyte Lysate

The mRNA encoding the β-subunit of the mitochondrial H+-ATP synthase (β-F1-ATPase) is localized in an approx. 150nm structure of the hepatocyte of mammals. In the present study, we have investigated the cis- and trans-acting factors involved in the generation of the ribonucleoprotein complex containing β-F1-ATPase mRNA. Two cis-acting elements (β1.2 and 3′β) have been identified. The β1.2 element is placed in the open reading frame, downstream of the region encoding the mitochondrial pre-sequence of the protein. The 3′β element is the 3′ non-translated region of the mRNA. Complex sets of proteins from the soluble and non-soluble fractions of the liver interact with the β1.2 and 3′β elements. A soluble p88, present also in reticulocyte lysate, displays binding specificity for both the cis-acting elements. Sedimentation and high-resolution in situ hybridization experiments showed that the structure containing the rat liver β-F1-ATPase mRNA is found in fractions of high sucrose concentration, where large polysomes sediment. Treatment of liver extracts with EDTA promoted the mobilization of β-F1-ATPase mRNA to fractions of lower sucrose concentration, suggesting that the structure containing β-F1-ATPase mRNA is a large polysome. Finally, in vitro reconstitution experiments with reticulocyte lysate, using either the full-length, mutant or chimaeric versions of β-F1-ATPase mRNA, reveal that the assembly of the β-F1-ATPase mRNA polysome requires the co-operation of both the cis-acting mRNA determinants. The present study illustrates the existence of an intramolecular RNA cross-talking required for the association of the mRNA with the translational machinery.

scholarly journals Characterization of single-chain antibody (sFv)-toxin fusion proteins produced in vitro in rabbit reticulocyte lysate.

1993 ◽  
Vol 268 (7) ◽  
pp. 5302-5308
Author(s):  
P.J. Nicholls ◽  
V.G. Johnson ◽  
S.M. Andrew ◽  
H.R. Hoogenboom ◽  
J.C. Raus ◽  
...  
Keyword(s):  
Fusion Proteins ◽  
Rabbit Reticulocyte Lysate ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Reticulocyte Lysate

INDUCTION OF SPECIFIC TRANSLATABLE MESSENGER RNA'S BY OESTROGEN AND PROGESTERONE

1972 ◽  
Vol 71 (2_Suppla) ◽  
pp. S381-S395 ◽  
Author(s):  
Bert W. O'Malley ◽  
Gary C. Rosenfeld ◽  
John P. Comstock ◽  
Anthony R. Means
Keyword(s):  
De Novo ◽  
Rabbit Reticulocyte Lysate ◽  
System Synthesis ◽  
Reticulocyte Lysate

ABSTRACT In this manuscript, we report the de novo synthesis of two specific chick proteins, ovalbumin and avidin, in a heterologous rabbit reticulocyte lysate in vitro system. Synthesis of these proteins is absolutely dependent on RNA extracted from oviduct and not RNA extracted from non-target tissues. Furthermore, we have demonstrated that the intracellular accumulations of ovalbumin mRNA and avidin mRNA are directly dependent upon prior stimulation by oestrogen and progesterone, respectively.

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