Investigation of the mode of nuclear control over protein synthesis in early development of loach and sea urchin

Development ◽  
1972 ◽  
Vol 28 (3) ◽  
pp. 491-509
Author(s):  
Maya R. Krigsgaber ◽  
A. A. Neyfakh

It is shown that in loach embryos the incorporation of precursors into protein takes place in the blastoderm cells only. The change of the rate of incorporation of labelled amino acids into protein of the blastoderm separated from the yolk at successive developmental stages reflects the changes in the level of protein synthesis in intact embryos of the same developmental stages. Typical periodic changes of the intensity of protein synthesis in early embryo-genesis of the loach are detected: low incorporation of amino acids at blastula stages is followed by an increase of synthesis during gastrulation and by a decrease with the onset of organogenesis. To study the genetic control over protein synthesis at various developmental stages of loach and sea-urchin embryos the effects of ionizing radiation and long-term treatment with actinomycin D have been examined. X-Irradiation doses produce an insignificant direct effect on protein synthesis, while radiation damage of the nuclear apparatus results in a gradual but ever increasing inhibition of protein synthesis. The inhibition of RNA synthesis with actinomycin or ionizing radiation damage of the nuclei produce essentially the same effect on the intensity of protein synthesis. Protein synthesis in androgenetic haploid hybrid embryos (loach ♀ × goldfish ♂) and in loach androgenetic haploid embryos after producing a partial elimination of chromosomes does not differ from ‘enucleated’ loach embryos completely deprived of chromosomes. These data suggest that high-polymeric RNA formed after the elimination of some chromsomes is unable to provide a normal level of protein synthesis. Protein synthesis is not controlled by the nuclei up to the stages of early blastula (sea urchin) and of late blastula (loach), being evidently programmed in oogenesis. To provide a rapid activation of protein synthesis in the course of gastrulation in the loach the function of the nuclei has to be realized during mid-blastula stages. An increase of the rate of the incorporation of amino acids at the stages of mesenchyme blastula in the sea urchin depends on the synthesis of RNA at the early blastula. At the same time protein synthesis during gastrulation in the loach and at mesenchyme blastula in the sea urchin is much less dependent on the simultaneous RNA synthesis. Protein synthesis at these stages seems to be provided by the long-living templates and controlled by non-gene mechanisms of the regulation of translation. Thus early embryonic differentiation in the loach and sea-urchin development is related to the activation of protein synthesis. The latter is provided by the preceding morphogenetic nuclear function, which makes protein synthesis relatively independent of simultaneous synthesis of templates that ensures subsequent development stages.

1970 ◽  
Vol 7 (2) ◽  
pp. 575-585
Author(s):  
J. B. GRIFFITHS

In a confluent culture of WI-38 cells the membrane area available for nutrient uptake is greatly reduced and the possibility exists that this reduction in uptake capacity of the cell is a contributory factor in contact inhibition. Insulin has been reported by many authors to facilitate glucose uptake and also to stimulate protein, DNA and RNA synthesis, glycolysis, pino-cytosis and growth in cultured cells. The effect of insulin on WI-38 cells was determined, therefore, to find out whether it enabled the cell to escape from contact inhibition of growth. The action of insulin was found to be dependent upon medium composition. Growth and protein synthesis were stimulated in Eagle's minimal essential medium, but not when this medium was supplemented with glucose and glutamine. Apparently insulin is only effective when high-energy compounds become limiting. Whilst insulin did not induce any post-confluent division, the protein content of cells was increased by 30%, and this was correlated with an increased rate of protein synthesis. Despite this increased activity in protein metabolism, the utilization of amino acids was less in the presence of insulin indicating that a control mechanism for more economical utilization of amino acids for protein synthesis was activated by insulin. Insulin had no effect on RNA synthesis, and only a slight inhibitory effect on DNA synthesis. Evidence was produced suggesting that insulin blocked cell division and encouraged differentiation. Glucose uptake and incorporation into the cell was stimulated by insulin, and this was especially noticeable after the cell sheet became confluent. The turnover of labelled glucose and derivatives was also enhanced by insulin and this was accompanied by a much higher rate of lactic acid production. It is concluded that insulin does not overcome contact inhibition and permit post-confluent division, but that it does enable the cell to take up and utilize nutrients more efficiently in confluent cultures with a resultant increase in metabolic activity and cell size.


1988 ◽  
Vol 8 (8) ◽  
pp. 3518-3525
Author(s):  
Z Y Gong ◽  
B P Brandhorst

An increased level of unpolymerized tubulin caused by depolymerization of microtubules in sea urchin larvae resulted in a rapid loss of tubulin mRNA, which was prevented by nearly complete inhibition of protein synthesis. Results of an RNA run-on assay indicated that inhibition of protein synthesis does not alter tubulin gene transcription. Analysis of the decay of tubulin mRNA in embryos in which RNA synthesis was inhibited by actinomycin D indicated that inhibition of protein synthesis prevents the destabilization of tubulin mRNA. The effect was similar whether mRNA was maintained on polysomes in the presence of emetine or anisomycin or displaced from the polysomes in the presence of puromycin or pactamycin; thus, the stabilization of tubulin mRNA is not dependent on the state of the polysomes after inhibition of protein synthesis. Even after tubulin mRNA declined to a low level after depolymerization of microtubules, it could be rescued by treatment of embryos with inhibitors of protein synthesis. Tubulin mRNA could be induced to accumulate prematurely in gastrulae but not in plutei if protein synthesis was inhibited, an observation that is indicative of the importance of the autogenous regulation of tubulin mRNA stability during embryogenesis. Possible explanations for the role of protein synthesis in the control of mRNA stability are discussed.


1988 ◽  
Vol 8 (8) ◽  
pp. 3518-3525 ◽  
Author(s):  
Z Y Gong ◽  
B P Brandhorst

An increased level of unpolymerized tubulin caused by depolymerization of microtubules in sea urchin larvae resulted in a rapid loss of tubulin mRNA, which was prevented by nearly complete inhibition of protein synthesis. Results of an RNA run-on assay indicated that inhibition of protein synthesis does not alter tubulin gene transcription. Analysis of the decay of tubulin mRNA in embryos in which RNA synthesis was inhibited by actinomycin D indicated that inhibition of protein synthesis prevents the destabilization of tubulin mRNA. The effect was similar whether mRNA was maintained on polysomes in the presence of emetine or anisomycin or displaced from the polysomes in the presence of puromycin or pactamycin; thus, the stabilization of tubulin mRNA is not dependent on the state of the polysomes after inhibition of protein synthesis. Even after tubulin mRNA declined to a low level after depolymerization of microtubules, it could be rescued by treatment of embryos with inhibitors of protein synthesis. Tubulin mRNA could be induced to accumulate prematurely in gastrulae but not in plutei if protein synthesis was inhibited, an observation that is indicative of the importance of the autogenous regulation of tubulin mRNA stability during embryogenesis. Possible explanations for the role of protein synthesis in the control of mRNA stability are discussed.


1974 ◽  
Vol 52 (6) ◽  
pp. 1178-1185 ◽  
Author(s):  
Joël de la Noüe

When fertilized eggs of Strongylocentrotus purpuratus, the purple sea urchin, are incubated in sea water containing 32PO43− and L-14C-valine, valine incorporation is inhibited in the presence of puromycin, cycloheximide, or chloramphenicol, but only the last inhibits the active transport of phosphate. Since chloramphenicol does not depress the egg respiration, it is likely that this drug acts specifically. This is in line with a lack of inhibitory effect of chloramphenicol on phosphate uptake at later developmental stages, except at the time of gastrulation, when the appearance of new phosphate carriers might well occur. It is suggested that the active transport carrier for phosphate, or some element required for its operation, is synthesized after fertilization, with the likely participation of the mitochondrial protein-synthesis machinery. A similar proposal holds for valine uptake.


2021 ◽  
Author(s):  
Geoffrey Henry Siwo

Life on earth relies on three types of information polymers- DNA, RNA and proteins. In all organisms and viruses, these molecules are synthesized by the copying of pre-existing templates. A triplet-based code known as the genetic code guides the synthesis of proteins by complex enzymatic machines that decode genetic information in RNA sequences. The origin of the genetic code is one of the most fundamental questions in biology. In this study, computational analysis of about 5,000 species level metagenomes using techniques for the analysis of human language suggests that the genomes of extant organisms contain relics of a distinct triplet code that potentially predates the genetic code. This code defines the relationship between adjacent triplets in DNA/RNA sequences, whereby these triplets predominantly differ by a single base. Furthermore, adjacent triplets encode amino acids that are thought to have emerged around the same period in the earth's early history. The results suggest that the order of triplets in primordial RNA sequences was associated with the availability of specific amino acids, perhaps due to a coupling of a triplet-based primordial RNA synthesis mechanism to a primitive mechanism of peptide bond formation. Together, this coupling could have given rise to early nucleic acid sequences and a system for encoding amino acid sequences in RNA, i.e. the genetic code. Thus, the central role of triplets in biology potentially extends to the primordial world, contributing to both the origins of genomes and the origins of genetically coded protein synthesis.


1976 ◽  
Vol 20 (3) ◽  
pp. 525-537
Author(s):  
J.M. Cameron ◽  
S.E. Hawkins

The injection of non-homologous cytoplasm into any strain of large free-living amoebae leads to a 60% inhibition of division amongst recipient cells. When the post-microsomal supernatant fraction of Amoeba discoides was injected into A. proteus, this inhibition of division was as high as 95%. The incorporation of tritiated precursors, either [3H]uridine or 3H-amino acids, into these inhibited amoebae was studied at various times after the injection of the inhibitory material using autoradiography. When cells were grown in [3H]uridine, autoradiographs indicated that RNA synthesis had ceased 2 days after the injection of non-homologous material. However, if [3H]uridine was injected into the inhibited cells, some synthesis of RNA could be detected up to 4 days after the injection of inhibitor. These results suggested that uptake of [3H]uridine was impaired and that one site of action of the inhibitory molecules was RNA synthesis for membrane components. Experiments with a variety of 3H-amino acids suggested that protein synthesis continued for at least 9 days after the injection of non-homologous cytoplasm, and that in these cells some informational RNA molecules were long-lived. There seemed to be accumulation of material containing [3H]lysine in the nuclei of control cells taken at random from cultures, and this was seen in the nuclei of inhibited cells 1 day after injection. However, 2 days after the injection of inhibitor, no accumulation of [3H]lysine-containing material was found in the nuclei.


Sign in / Sign up

Export Citation Format

Share Document