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Biomedical subjects

S Ochoa

Publications and source records attributed to S Ochoa.

At least 91 records · Page 5Linked to original sources
PubMed1974

Effect of development on the translation of messenger RNA in Artemia salina embryos.

Cell-free preparations from encapsulated Artemia salina embryos readily translate poly(U), but there is no endogenous protein synthesis or translation of added natural mRNA until development resumes upon incubation in saline. High-salt-washed 80S ribosomes and highspeed supernatant (cytosol) were prepared from undeveloped eggs and from eggs allowed to develop to a stage prior to hatching. Endogenous protein synthesis occurs only with ribosomes from developed embryos, whether with undeveloped or developed cytosol. This is mainly elongation of preformed polypeptide chains, for it is largely resistant to edeine, an inhibitor of chain initiation. Edeinesensitive translation of added natural mRNA occurs with both developed and undeveloped ribosomes but requires developed cytosol. Translation of brome mosaic virus RNA is not much, if at all, further stimulated by high-saltwash of developed ribosomes, but that of globin mRNA is markedly enhanced. Levels of the chain initiation factor EIF-1 are the same before and after development. These results are consistent with the view that resumption of developemtn is triggered by transcription, with ensuing translation of the resulting messengers to yield, among other proteins, mRNA-recognizing initiation factors of the IF-3 type which are partly free in the cytosol and partly ribosome-bound. The data also suggest that different factors may be involved in the translation of brome mosaic virus RNA and globin mRNA by this system.

Animals↗
PubMed1974

Inhibition of polypeptide chain initiation in Escherichia coli by elongation factor G.

We have previously reported the isolation from E. coli of a specific inhibitor of polypeptide chain initiation that is rendered ineffective when active aminoacylation of transfer RNA is taking place; this is normally the case during natural messenger RNA translation. Surprisingly, the inhibitory activity appears to be a hitherto unrecognized property of the chain elongation factor G. The following hold for preparations purified for either translocase or inhibitor activity: (1) equal electrophoretic mobility on polyacrylamide gels; (2) equal specific activities for (a) inhibition of initiation, (b) translocation, and (c) ribosome-dependent, uncoupled GTPase; and (3) similar heat sensitivity of translocase and inhibitor activities in a temperature-sensitive E. coli mutant with an altered elongation factor G. Different sites are apparently involved in translocation and inhibition because the former, but not the latter, is sensitive to p-chloromercuribenzoate and fusidic acid.

Acylation↗
PubMed1973

A specific inhibitor of polypeptide-chain initiation in Escherichia coli.

An inhibitor of polypeptide-chain initiation was isolated from E. coli cells. This protein inhibits formation of the 30S or 70S initiation complex with either fMet-tRNA(f) as initiator and AUG, MS2 RNA, or late T4 RNA as messenger, or acPhe-tRNA as initiator and poly(U) as messenger. Chain elongation, e.g., poly(U) translation at high Mg(2+) concentration, is not inhibited. The inhibitor is rendered ineffective when active aminoacylation of tRNA is taking place, e.g., during natural mRNA translation. This inhibitor is distinct from the so-called interference (i) factors, which interfere exclusively with the action of initiation factor 3. Since the new inhibitor can apparently be turned on and off, it may have a regulatory function in translation.

Bacterial Proteins↗
PubMed1973

Conformational control of the interaction of eukaryotic elongation factors EF-1 and EF-2 with ribosomes.

As in the case with prokaryotic systems, Artemia salina elongation factors EF-1 and EF-2 interact with a common site or with closely overlapping sites on the Artemia ribosome. This feature of ribosomal design must restrict interaction with the ribosome to only one of the factors at alternating steps of chain elongation. In support of this view we find that EF-1, but not EF-2, interacts with the post-translocation ribosome, whereas the reverse is true of the pre-translocation ribosome. Conformational changes probably account for the alternating selectivity of the translating ribosome for each elongation factor.

Animals↗
PubMed1972

Polypeptide chain initiation in eukaryotes: functional identity of supernatant factor from various sources.

Eukaryotic cells contain polypeptide chain initiation factors that, like the prokaryotic initiation factor IF-2, promote the AUG-dependent binding of fMet-tRNA(f) to the small ribosomal subunit. The bound amino-acyl-tRNA is directly convertible to fMet-puromycin upon addition of 60S subunit. The reaction is sensitive to initiation inhibitors such as aurintricarboxylic acid and edeine but, unlike its prokaryotic counterpart, it does not require GTP. Factors that catalyze the binding and fMet-puromycin reactions with ribosomal subunits from Artemia salina embryos are present in postribosomal supernatants of Artemia, mouse fibroblasts (L cells), and rat liver, as well as in salt washes of rabbit reticulocyte ribosomes. However, whereas all three supernatant factors, like Escherichia coli IF-2, are sensitive to SH-binding reagents, the reticulocyte factor is not. The rat liver and Artemia factors function indiscriminately with Artemia or rat liver ribosomes, but the Artemia factor and E. coli initiation factor IF-2 are not interchangeable.

Animals↗
PubMed1972

Polypeptide chain initiation and stepwise elongation with Artemia ribosomes and factors.

The supernatant initiation factor from Artemia salina embryos promotes, besides the AUG-dependent binding of fMet-tRNA(f), the poly(U)-dependent binding of N-acetylPhe-tRNA to 40S ribosomal subunits; the bound N-acylaminoacyl-tRNA reacts directly with puromycin upon addition of 60S subunits. Both the binding reaction and the synthesis of N-acylaminoacyl-puromycin occur in the absence of GTP or other ribonucleoside triphosphates. To a smaller extent, the factor also mediates the 40S ribosomal binding of Met-tRNA(f) and Phe-tRNA; in this case, the bound aminoacyl-tRNA is less reactive with puromycin. After the poly(U)- and supernatant factor-dependent binding of N-acetylPhe-tRNA to 40S subunits at low Mg(2+) concentration, binding of a second aminoacyl-tRNA (Phe-tRNA), with ensuing formation of the first peptide bond, is dependent upon the addition of the 60S subunit, elongation factor EF-1, and GTP. Further growth of the polypeptide chain requires translocation and is, therefore, dependent upon the addition of elongation factor EF-2. As with the Escherichia coli system, once requirements for translation of the third codon have been met, no further additions are necessary for elongation of a peptide chain.

Acetates↗
PubMed1971

A supernatant factor involved in initiation complex formation with eukaryotic ribosomes.

Embryos of the brine shrimp, Artemia salina, were used in a study of polypeptide chain initiation in an in vitro system from a eukaryote. A protein, isolated from the high-speed supernatant, has been highly purified and shown to have properties that suggest it is the eukaryotic equivalent of the Escherichia coli initiation factor F(2): It promotes the AUG-dependent binding of fMet-tRNA (E. coli) to the Artemia 40S ribosomal subunit, but not to either the 60S or 80S species; the bound fMet-tRNA is placed in a site on the smaller subunit from which it reacts with puromycin upon addition of the 60S subunit; and the activity is sensitive to aurintricarboxylic acid and edeine, specific inhibitors of initiation. The factor, a basic protein of molecular weight about 100,000, is inactivated by N-ethylmaleimide, an SH-binding reagent, and is clearly distinct from the Artemia elongation factors, T(1) and T(2). In addition, the factor stimulates the poly(U)-dependent binding of Phe-tRNA (E. coli) to the Artemia 40S ribosomal subunit. This reaction, though similar to the fMet-tRNA-binding reaction, differs in that the bound Phe-tRNA is largely resistant to release by puromycin.

Adenine Nucleotides↗