Dataset Information

The dissociation of Yoshida hepatoma ribosomes into active particles.

ABSTRACT: 1. The sedimentation pattern of Yoshida hepatoma ribosomes shows mainly a high dimer peak and an intermediate peak between those of monomer and dimer. 2. The treatment of the postmitochondrial supernatant with EDTA or potassium chloride leads to the dissociation of ribosomes into subunits, through a decrease ofthe ribosomal Mg(2+)/phosphorus ratio. Provided that the deprivation of endogenous Mg(2+) is not complete, the subunits reassociate into active polyribosomes after addition of magnesium chloride to the medium. If the Mg(2+)/phosphorus ratio is lowered below 0.01 (mumol/mumol), structural changes occur, that become evident by a loss of protein and by a decreased sedimentation rate, which render the subribosomal particles unable to reassociate. 3. In the re-formation of polyribosomes from subunits, during the progressive increase of the concentration of magnesium chloride in the medium, the formation of monomeric ribosomes seems to be intermediate. 4. A dimerization of monomers and subunits occurs at concentrations of magnesium chloride greater than those required for the re-formation of polyribosomes and a preincubation for 1h at 0 degrees C is necessary for the maximum dimerization, whereas the complete reconstitution of polyribosomes is immediate.

SUBMITTER: Gravela E

PROVIDER: S-EPMC1176497 | biostudies-other | 1971 Jan

REPOSITORIES: biostudies-other

ACCESS DATA

Similar Datasets

Project description:1. The ability of Escherichia coli ribosomes to function in poly(U)-directed protein synthesis was measured at elevated temperatures by using thermostable supernatant factors from Bacillus stearothermophilus. The amount of polyphenylalanine synthesized at 55 degrees C was about the same as at 37 degrees C, but the rate of synthesis was increased approximately fivefold. At 60 degrees C the activity of the ribosomes was halved. 2. E. coli ribosomes can sustain the loss of approx. 10% of the double-helical secondary structure of RNA without losing activity. 3. Within the active ribosome the double-helical secondary structure of the rRNA moiety is stabilized compared with isolated rRNA, as judged by enzymic hydrolysis and by measurements of E(260). 4. The main products, over the range 0-55 degrees C, of ribonuclease T(1) digestion of the smaller subribosomal particle of E. coli were two fragments (s(0) (20,w) 15S and 25.3S) of approximately one-quarter and three-quarters of the size of the intact molecule, revealing the presence of a ;weak spot' where intramolecular bonds appear insufficient to hold the fragments together. 5. Subribosomal particles of B. stearothermophilus were more stable to heating, by approx. 10 degrees C, than those of E. coli, and the stabilization of double-helical secondary structure of the RNA moiety was more striking. 6. Rabbit reticulocyte ribosomes were active in poly(U)-directed protein synthesis at 45 degrees C, and half the activity was lost after heating to 53 degrees C. Active subribosomal particles of rabbit reticulocytes and of oocytes of Xenopus laevis, like the bacterial subribosomal particles, underwent a conformational change to a slower-sedimenting form on heating. The temperature range of the transition depended on the species. 7. Slower-sedimenting particles, whether produced by EDTA treatment or by heating, had different ;melting' profiles compared with active subribosomal particles, providing another indication of conformational differences. 8. Comparison of the properties of the various subribosomal particles revealed greater variation in the secondary structure of the protein moieties (judged by measurement of circular dichroism) than in the secondary structure of the RNA moieties, which appeared to have features in common.

Dataset Information

The dissociation of Yoshida hepatoma ribosomes into active particles.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets