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. 1972 May;69(5):1146–1150. doi: 10.1073/pnas.69.5.1146

Ovalbumin Messenger RNA of Chick Oviduct: Partial Characterization, Estrogen Dependence, and Translation In Vitro

A R Means 1,2,3,4, J P Comstock 1,2,3,4, G C Rosenfeld 1,2,3,4, B W O'Malley 1,2,3,4
PMCID: PMC426650  PMID: 4504328

Abstract

A rapidly-labeled RNA fraction can be isolated from hen oviduct polysomes that has characteristics of the messenger RNA (mRNA) for the cell-specific protein, ovalbumin. This RNA, which sediments in the 8-17S region of sucrose gradients, possesses properties suggestive of the presence of a polyadenylic acid sequence and can be translated with fidelity in a cell-free protein synthesizing system derived from rabbit reticulocytes. The identity of the protein product as ovalbumin is confirmed by three methods, and translation of ovalbumin mRNA is shown to be dependent both on amount of exogenous mRNA and incubation time. Both rate and extent of ovalbumin synthesis is enhanced by the addition of a protein extract from ribosomes that contains peptide chain initiation factors. Finally, the presence of this specific mRNA is shown to be estrogen-dependent: it is induced by estrogen administration to immature chicks, disappears upon cessation of estrogen treatment, and can be reinduced by a single injection of estrogen to chicks that have been pretreated with estrogen and then withdrawn from the hormone.

Keywords: poly(A) sequence, estrogen action, protein synthesis, sucrose gradient centrifugation

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burr H., Lingrel J. B. Poly A sequences at the 3' termini of rabbit globin mRNAs. Nat New Biol. 1971 Sep 8;233(36):41–43. doi: 10.1038/newbio233041a0. [DOI] [PubMed] [Google Scholar]
  2. Darnell J. E., Philipson L., Wall R., Adesnik M. Polyadenylic acid sequences: role in conversion of nuclear RNA into messenger RNA. Science. 1971 Oct 29;174(4008):507–510. doi: 10.1126/science.174.4008.507. [DOI] [PubMed] [Google Scholar]
  3. Darnell J. E., Wall R., Tushinski R. J. An adenylic acid-rich sequence in messenger RNA of HeLa cells and its possible relationship to reiterated sites in DNA. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1321–1325. doi: 10.1073/pnas.68.6.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gilbert J. M., Anderson W. F. Cell-free hemoglobin synthesis. II. Characteristics of the transfer ribonucleic acid-dependent assay system. J Biol Chem. 1970 May 10;245(9):2342–2349. [PubMed] [Google Scholar]
  5. Kohler P. O., Grimley P. M., O'Malley B. W. Estrogen-induced cytodifferentiation of the ovalbumin-secreting glands of the chick oviduct. J Cell Biol. 1969 Jan;40(1):8–27. doi: 10.1083/jcb.40.1.8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lee S. Y., Mendecki J., Brawerman G. A polynucleotide segment rich in adenylic acid in the rapidly-labeled polyribosomal RNA component of mouse sarcoma 180 ascites cells. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1331–1335. doi: 10.1073/pnas.68.6.1331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lockard R. E., Lingrel J. B. The synthesis of mouse hemoglobin beta-chains in a rabbit reticulocyte cell-free system programmed with mouse reticulocyte 9S RNA. Biochem Biophys Res Commun. 1969 Oct 8;37(2):204–212. doi: 10.1016/0006-291x(69)90720-7. [DOI] [PubMed] [Google Scholar]
  8. Means A. R., Abrass I. B., O'Malley B. W. Protein biosynthesis on chick oviduct polyribosomes. I. Changes during estrogen-mediated tissue differentiation. Biochemistry. 1971 Apr 27;10(9):1561–1570. doi: 10.1021/bi00785a009. [DOI] [PubMed] [Google Scholar]
  9. O'Malley B. W., McGuire W. L., Kohler P. O., Korenman S. G. Studies on the mechanism of steroid hormone regulation of synthesis of specific proteins. Recent Prog Horm Res. 1969;25:105–160. doi: 10.1016/b978-0-12-571125-8.50006-5. [DOI] [PubMed] [Google Scholar]
  10. O'Malley B. W., McGuire W. L., Korenman S. G. Estrogen stimulation of synthesis of specific proteins and RNA polymerase activity in the immature chick oviduct. Biochim Biophys Acta. 1967 Aug 22;145(1):204–207. doi: 10.1016/0005-2787(67)90679-x. [DOI] [PubMed] [Google Scholar]
  11. O'Malley B. W., McGuire W. L. Studies on the mechanism of estrogen-mediated tissue differentiation: regulation of nuclear transcription and induction of new RNA species. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1527–1534. doi: 10.1073/pnas.60.4.1527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Oka T., Schimke R. T. Interaction of estrogen and progesterone in chick oviduct development. II. Effects of estrogen and progesterone on tubular gland cell function. J Cell Biol. 1969 Oct;43(1):123–137. doi: 10.1083/jcb.43.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. RHODES M. B., AZARI P. R., FEENEY R. E. Analysis, fractionation, and purification of egg white proteins with cellulose-cation exchanger. J Biol Chem. 1958 Jan;230(1):399–408. [PubMed] [Google Scholar]
  14. Rhoads R. E., McKnight G. S., Schimke R. T. Synthesis of ovalbumin in a rabbit reticulocyte cell-free system programmed with hen oviduct ribonucleic acid. J Biol Chem. 1971 Dec 10;246(23):7407–7410. [PubMed] [Google Scholar]
  15. Rosenfeld G. C., Comstock J. P., Means A. R., O'Malley B. W. Estrogen-induced synthesis of ovalbumin messenger RNA and its translation in a cell-free system. Biochem Biophys Res Commun. 1972 Feb 25;46(4):1695–1703. doi: 10.1016/0006-291x(72)90805-4. [DOI] [PubMed] [Google Scholar]
  16. Ross J., Aviv H., Scolnick E., Leder P. In vitro synthesis of DNA complementary to purified rabbit globin mRNA (RNA-dependent DNA polymerase-reticulocyte-hemoglobin-density gradient centrifugation-oligo(dT) primer). Proc Natl Acad Sci U S A. 1972 Jan;69(1):264–268. doi: 10.1073/pnas.69.1.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stavnezer J., Huang R. C. Synthesis of a mouse immunoglobulin light chain in a rabbit reticulocyte cell-free system. Nat New Biol. 1971 Apr 7;230(14):172–176. doi: 10.1038/newbio230172a0. [DOI] [PubMed] [Google Scholar]
  18. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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