Translation of ovalbumin mRNA in Xenopus laevis oocytes. Characterization of the system and effects of estrogen on injected mRNA populations

L. Chan, Peter Kohler, B. W. O'Malley

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17 Citations (Scopus)

Abstract

Ovalbumin messenger RNA (mRNA(ov)) purified from hen oviduct was injected into Xenopus laevis oocytes. The oocytes were incubated in culture medium containing [3H] leucine. Analysis of the oocyte cytosol on Sephadex G 150 columns demonstrated a peak of radioactivity which cochromatographed with authentic ovalbumin. Radioactive protein contained in this peak was precipitated by ovalbumin antiserum, co electrophoresed with ovalbumin on sodium dodecyl sulfate (SDS) and urea gels at pH 8.7, and eluted with the protein at the same pH (4.8) on CM cellulose chromatography. Injection of increasing amounts of mRNA(ov) was found to elicit a linear response in terms of ovalbumin synthesis. Moreover, there was linear incorporation of radioactivity into microinjected oocytes over a minimum period of 91 h. Less than 1 ng mRNA(ov) was detected in this system. Ovalbumin mRNA activity was present in RNA preparations from chicks treated with estrogen but was undetectable in animals withdrawn from the hormone. This study constitutes an initial demonstration of a steroid hormone induced alteration in mRNA population as assayed in intact viable heterologous cells.

Original languageEnglish (US)
Pages (from-to)576-585
Number of pages10
JournalJournal of Clinical Investigation
Volume57
Issue number3
StatePublished - 1976
Externally publishedYes

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Ovalbumin
Xenopus laevis
Protein Biosynthesis
Oocytes
Estrogens
Messenger RNA
Population
Radioactivity
Hormones
Oviducts
Leucine
Cellulose
Sodium Dodecyl Sulfate
Cytosol
Culture Media
Urea
Chromatography
Immune Sera
Proteins
Gels

ASJC Scopus subject areas

  • Medicine(all)

Cite this

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title = "Translation of ovalbumin mRNA in Xenopus laevis oocytes. Characterization of the system and effects of estrogen on injected mRNA populations",
abstract = "Ovalbumin messenger RNA (mRNA(ov)) purified from hen oviduct was injected into Xenopus laevis oocytes. The oocytes were incubated in culture medium containing [3H] leucine. Analysis of the oocyte cytosol on Sephadex G 150 columns demonstrated a peak of radioactivity which cochromatographed with authentic ovalbumin. Radioactive protein contained in this peak was precipitated by ovalbumin antiserum, co electrophoresed with ovalbumin on sodium dodecyl sulfate (SDS) and urea gels at pH 8.7, and eluted with the protein at the same pH (4.8) on CM cellulose chromatography. Injection of increasing amounts of mRNA(ov) was found to elicit a linear response in terms of ovalbumin synthesis. Moreover, there was linear incorporation of radioactivity into microinjected oocytes over a minimum period of 91 h. Less than 1 ng mRNA(ov) was detected in this system. Ovalbumin mRNA activity was present in RNA preparations from chicks treated with estrogen but was undetectable in animals withdrawn from the hormone. This study constitutes an initial demonstration of a steroid hormone induced alteration in mRNA population as assayed in intact viable heterologous cells.",
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T1 - Translation of ovalbumin mRNA in Xenopus laevis oocytes. Characterization of the system and effects of estrogen on injected mRNA populations

AU - Chan, L.

AU - Kohler, Peter

AU - O'Malley, B. W.

PY - 1976

Y1 - 1976

N2 - Ovalbumin messenger RNA (mRNA(ov)) purified from hen oviduct was injected into Xenopus laevis oocytes. The oocytes were incubated in culture medium containing [3H] leucine. Analysis of the oocyte cytosol on Sephadex G 150 columns demonstrated a peak of radioactivity which cochromatographed with authentic ovalbumin. Radioactive protein contained in this peak was precipitated by ovalbumin antiserum, co electrophoresed with ovalbumin on sodium dodecyl sulfate (SDS) and urea gels at pH 8.7, and eluted with the protein at the same pH (4.8) on CM cellulose chromatography. Injection of increasing amounts of mRNA(ov) was found to elicit a linear response in terms of ovalbumin synthesis. Moreover, there was linear incorporation of radioactivity into microinjected oocytes over a minimum period of 91 h. Less than 1 ng mRNA(ov) was detected in this system. Ovalbumin mRNA activity was present in RNA preparations from chicks treated with estrogen but was undetectable in animals withdrawn from the hormone. This study constitutes an initial demonstration of a steroid hormone induced alteration in mRNA population as assayed in intact viable heterologous cells.

AB - Ovalbumin messenger RNA (mRNA(ov)) purified from hen oviduct was injected into Xenopus laevis oocytes. The oocytes were incubated in culture medium containing [3H] leucine. Analysis of the oocyte cytosol on Sephadex G 150 columns demonstrated a peak of radioactivity which cochromatographed with authentic ovalbumin. Radioactive protein contained in this peak was precipitated by ovalbumin antiserum, co electrophoresed with ovalbumin on sodium dodecyl sulfate (SDS) and urea gels at pH 8.7, and eluted with the protein at the same pH (4.8) on CM cellulose chromatography. Injection of increasing amounts of mRNA(ov) was found to elicit a linear response in terms of ovalbumin synthesis. Moreover, there was linear incorporation of radioactivity into microinjected oocytes over a minimum period of 91 h. Less than 1 ng mRNA(ov) was detected in this system. Ovalbumin mRNA activity was present in RNA preparations from chicks treated with estrogen but was undetectable in animals withdrawn from the hormone. This study constitutes an initial demonstration of a steroid hormone induced alteration in mRNA population as assayed in intact viable heterologous cells.

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