Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Biochem J
1989 Mar 01;2582:553-61. doi: 10.1042/bj2580553.
Show Gene links
Show Anatomy links
Characterization of translation systems in vitro from three developmental stages of Strongylocentrotus purpuratus.
Lopo AC
,
Lashbrook CC
,
Hershey JW
.
???displayArticle.abstract???
We have developed and characterized cell-free systems active in translation from unfertilized eggs, 30-min zygotes and hatched blastulae of the sea urchin Strongylocentrotus purpuratus. The ion concentrations selected for preparation of the lysates were 150 mM-K+, 40 mM-Na+, 40 mM-Cl-, 5 x 10(-7) M free Ca2+ and 1 mM free Mg2+. It was necessary to include the ribonuclease inhibitor RNas in the preparations to obtain full activity consistently. The pH optimum was 7.2 and was extremely sharp for the three S. purpuratus lysates. The temperature optima of the three lysates were remarkably similar to those of the intact unfertilized egg and embryos. Lysates from unfertilized egg and 30-min zygotes showed a temperature optimum at 15 degrees C. The hatched blastula lysate showed a broader temperature optimum with a shift to about 20 degrees C. The optimized lysates incorporated radiolabelled amino acids into polypeptides for up to 90 min. The polypeptides synthesized ranged in Mr from 200,000 to 20,000, suggesting that the mRNA in the lysates was intact and capable of directing the synthesis of complete polypeptides. Furthermore, the three lysates were capable of initiation, as demonstrated by inhibition of initiation using the inhibitors edeine and 7-methylguanosine 5''-triphosphate (m7GTP). At 15 degrees C, the transit times for the three lysates were: unfertilized egg, 40 min; 30-min zygotes and hatched blastula lysates, 20 min. These transit times are similar to those of intact eggs and embryos, and significantly, reflect the two-fold increase in elongation rate seen following fertilization in intact embryos. Thus, these lysates display many features and characteristic responses typical of intact eggs and embryos, indicating that the lysates should be useful tools for the analysis of translation control in early embryogenesis.
Bradford,
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
1976, Pubmed
Bradford,
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
1976,
Pubmed
Brandhorst,
Two-dimensional gel patterns of protein synthesis before and after fertilization of sea urchin eggs.
1976,
Pubmed
,
Echinobase
Brandis,
Translation of oogenetic mRNA in sea urchin eggs and early embryos. Demonstration of a change in translational efficiency following fertilization.
1978,
Pubmed
,
Echinobase
Brandis,
Elevation of protein synthesis is a complex response to fertilisation.
1979,
Pubmed
,
Echinobase
Brostrom,
Identification of a Ca2+ requirement for protein synthesis in eukaryotic cells.
1983,
Pubmed
Davidson,
Molecular biology of the sea urchin embryo.
1982,
Pubmed
,
Echinobase
Eisenstein,
Characterization of a protein synthesis system from rat liver. Translation of endogenous and exogenous messenger RNA.
1984,
Pubmed
Epel,
Protein synthesis in sea urchin eggs: a "late" response to fertilization.
1967,
Pubmed
,
Echinobase
Epel,
An analysis of the partial metabolic derepression of sea urchin eggs by ammonia: the existence of independent pathways.
1974,
Pubmed
,
Echinobase
Fan,
Regulation of protein synthesis in mammalian cells. II. Inhibition of protein synthesis at the level of initiation during mitosis.
1970,
Pubmed
Foerder,
Release of ovoperoxidase from sea urchin eggs hardens the fertilization membrane with tyrosine crosslinks.
1977,
Pubmed
,
Echinobase
Gambino,
Properties of the ribosomal salt wash from unfertilized and fertilized sea urchin eggs and its effect on natural mRNA translation.
1973,
Pubmed
,
Echinobase
Goustin,
Protein synthesis, polyribosomes, and peptide elongation in early development of Strongylocentrotus purpuratus.
1981,
Pubmed
,
Echinobase
Hansen,
Inhibitor of translational initiation in sea urchin eggs prevents mRNA utilization.
1987,
Pubmed
,
Echinobase
Hille,
Efficiency of protein synthesis after fertilisation of sea urchin eggs.
1979,
Pubmed
,
Echinobase
Humphreys,
Measurements of messenger RNA entering polysomes upon fertilization of sea urchin eggs.
1971,
Pubmed
,
Echinobase
Humphreys,
Efficiency of translation of messenger-RNA before and after fertilization in sea urchins.
1969,
Pubmed
,
Echinobase
Ilan,
Translation of maternal messenger ribonucleoprotein particles from sea urchin in a cell-free system from unfertilized eggs and product analysis.
1978,
Pubmed
,
Echinobase
Jackson,
The roles of sugar phosphates and thiol-reducing systems in the control of reticulocyte protein synthesis.
1983,
Pubmed
Laemmli,
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
1970,
Pubmed
Lopo,
Translational control in early sea urchin embryogenesis: initiation factor eIF4F stimulates protein synthesis in lysates from unfertilized eggs of Strongylocentrotus purpuratus.
1988,
Pubmed
,
Echinobase
Lopo,
Translational initiation factors from sea urchin eggs and embryos: functional properties are highly conserved.
1986,
Pubmed
,
Echinobase
Mandley,
Putative nuclease-sensitive control element in unfertilized eggs of the sea urchin Lytechinus pictus.
1987,
Pubmed
,
Echinobase
Nielsen,
Evidence for control of protein synthesis in HeLa cells via the elongation rate.
1980,
Pubmed
Regier,
Absolute rates of protein synthesis in sea urchins with specific activity measurements of radioactive leucine and leucyl-tRNA.
1977,
Pubmed
,
Echinobase
Shen,
Direct measurement of intracellular pH during metabolic derepression of the sea urchin egg.
1978,
Pubmed
,
Echinobase
Steinhardt,
Intracellular calcium release at fertilization in the sea urchin egg.
1977,
Pubmed
,
Echinobase
Vacquier,
Methods for quantitating sea urchin sperm-egg binding.
1973,
Pubmed
,
Echinobase
Whitaker,
Ionic regulation of egg activation.
1982,
Pubmed
,
Echinobase
Winkler,
Multiple levels of regulation of protein synthesis at fertilization in sea urchin eggs.
1985,
Pubmed
,
Echinobase
Winkler,
The cytoplasmic 4 S translation inhibitory RNA species of chick embryonic muscle. Effect on mRNA binding to 43 S initiation complex.
1983,
Pubmed
Winkler,
Dual ionic controls for the activation of protein synthesis at fertilization.
1980,
Pubmed
,
Echinobase
Yablonka-Reuveni,
Isolation and distribution of elongation factor 2 in eggs and embryos of sea urchins.
1983,
Pubmed
,
Echinobase
Zucker,
Stimulation of globin-chain initiation by hemin in the reticulocyte cell-free system.
1968,
Pubmed
