Dissociation of yeast ribosomes; The effects of hydrostatic pressure and KCl

Amesz, W.J.C.; Bevers, Eduard M.; Bloemers, H. P. J.

doi:10.1007/bf00357403

Cited by 4 publications

References 15 publications

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The puromycin reaction mediated by yeast ribosomes in high salt

Mast

Bloemers

1973

Molecular Biology Reports

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The extend of the reaction between puromycin and yeast peptidyl-tRNA prelabeled in vitro was determined by measuring the distribution of trichloroacetic acid precipitable material in isokinetic sucrose gradients in the presence of 0.5 M KCl.Thus it was found that increasing amounts of puromycin remove increasing amounts of peptidyl-tRNA from the 80S position in the gradient. The extend of the reaction, however, was independent of pretreatment of the ribosomes with inhibitors of the translocation indicating that peptidyl-tRNA at the donor and at the acceptor site of the ribosomes are equally accessible to puromycin at 0.5 M KCl.The exposure of both ribosomal binding sites to puromycin in high salt is accompanied by an enhanced reactivity of puromycin towards peptidyl-tRNA. The ED50 determined by measuring the inhibition by puromycin of the poly-U dependent phenylalanine incorporation drops from 5×10(-5) M below 250 mM KCl to 5×10(-6) M at 300 mM and higher concentrations of KCl.

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The puromycin reaction mediated by yeast ribosomes in high salt

Mast

Bloemers

1973

Molecular Biology Reports

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Dissociability of free and peptidyl-tRNA bound ribosomes

Surguchov¹,

Fominykch²,

Lyzlova³

1978

Mol Biol Rep

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The influence of peptidyl-tRNA on the dissociation of yeast 80 S ribosomes into subunits was studied. For this purpose temperature-sensitive (ts) suppressor strain of yeast Saccharomyces cervisiae carrying a defect in peptide chain termination was used. It was found that peptidyl-tRNA did not influence the dissociation of ribosomes either at high salt concentration or in the presence of dissociation factor (DF) from yeast. After dissociation of yeast ribosomes in 0.5 M KCl, peptidyl-tRNA remains bound to the 60 S subunit. Some characteristics of the termination process and release of nascent polypeptides from yeast ribosomes are discussed.

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Ribosomal Subunits From Neonatal Mouse Brain Highly Active in Polyphenylalanine Synthesis

Saag

Greeff

1979

Journal of Neurochemistry

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Abstract— A highly active subcellular protein synthesising system is described, in which uncomplexed ribosomes isolated from 5 to 7 day old mouse brain can be reprogrammed with polyuridylic acid. Either purified free polyribosomes or microsomes were used as the starting material for the preparation of uncomplexed ribosomes by treatment with 0.5m‐KCl and puromycin. After reduction of the salt concentration 80S ribosomes were isolated by washing through sucrose. When, subsequently, zonal centrifugation in equivolumetric sucrose gradients containing 0.5 m‐KCI was performed, purified ribosomal subunits were obtained. Cross‐contamination of subunits was less than 5%. Re‐associated ribosomes and recombined isolated ribosomal subunits both showed high activities in vitro. Incorporation levels of 50–60 phenylalanine residues per ribosome could be reached, at a rate of 0.5–2.0 residues/min/ribosome, depending on the activity of the high speed supernatant enzymes added. It was shown by paper chromatography of the cell‐free product that only oligophenylalanine formation takes place. It was estimated that 6&70% of the ribosomes present in vitro were actively participating in the protein synthesis process.

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Dissociation of yeast ribosomes; The effects of hydrostatic pressure and KCl

Cited by 4 publications

References 15 publications

The puromycin reaction mediated by yeast ribosomes in high salt

The puromycin reaction mediated by yeast ribosomes in high salt

Dissociability of free and peptidyl-tRNA bound ribosomes

Ribosomal Subunits From Neonatal Mouse Brain Highly Active in Polyphenylalanine Synthesis

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