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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1973 Feb;70(2):406–411. doi: 10.1073/pnas.70.2.406

Cytoplasmic Adenylation and Processing of Maternal RNA

Isabel Slater *, David Gillespie †,, D W Slater *
PMCID: PMC433270  PMID: 4510284

Abstract

Molecular hybridization between [3H]-poly(U) and unlabeled RNA prepared from sea urchin eggs and embryos has been used to contrast the subcellular localization as well as the size distribution of adenylylated maternal RNA preexisting in the unfertilized egg with that adenylylated as a function of fertilization. Evidence reported establishes that such preadenylylated genetic messages are predominantly located in the ovum's subribosomal fraction and that fertilization elicits a rapid reallocation of these latent transcripts into the zygote's ribosomal fraction. Examination of the size distribution of the adenylylated RNA further demonstrates that the unfertilized egg contains a substantial population of RNA transcripts of exceptionally high molecular weight that are used as primers for the 2-fold net synthesis of poly(A) that follows fertilization. The poly(A)-rich tracts are shown to be covalently bonded to RNA. Assessment of the poly(A) content of nuclear and cytoplasmic fractions suggests that the function of poly(A) is not confined to the transport of genetic messages from the nucleus.

Keywords: sea urchin eggs and embryos, poly(A), subcellular localization, size distribution, covalent bonding

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Selected References

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

  1. Adesnik M., Darnell J. E. Biogenesis and characterization of histone messenger RNA in HeLa cells. J Mol Biol. 1972 Jun 28;67(3):397–406. doi: 10.1016/0022-2836(72)90458-5. [DOI] [PubMed] [Google Scholar]
  2. Armstrong J. A., Edmonds M., Nakazato H., Phillips B. A., Vaughn M. H. Polyadenylic acid sequences in the virion RNA of poliovirus and Eastern Equine Encephalitis virus. Science. 1972 May 5;176(4034):526–528. doi: 10.1126/science.176.4034.526. [DOI] [PubMed] [Google Scholar]
  3. Brawerman G., Mendecki J., Lee S. Y. A procedure for the isolation of mammalian messenger ribonucleic acid. Biochemistry. 1972 Feb 15;11(4):637–641. doi: 10.1021/bi00754a027. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Chamberlain J. P. RNA synthesis in anucleate egg fragments and normal embryos of the sea urchin, Arbacia punctulata. Biochim Biophys Acta. 1970 Jul 16;213(1):183–193. doi: 10.1016/0005-2787(70)90019-5. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Davidson E. H., Hough B. R. Genetic information in oocyte RNA. J Mol Biol. 1971 Mar 28;56(3):491–506. doi: 10.1016/0022-2836(71)90396-2. [DOI] [PubMed] [Google Scholar]
  9. Edmonds M., Caramela M. G. The isolation and characterization of adenosine monophosphate-rich polynucleotides synthesized by Ehrlich ascites cells. J Biol Chem. 1969 Mar 10;244(5):1314–1324. [PubMed] [Google Scholar]
  10. Edmonds M., Vaughan M. H., Jr, Nakazato H. Polyadenylic acid sequences in the heterogeneous nuclear RNA and rapidly-labeled polyribosomal RNA of HeLa cells: possible evidence for a precursor relationship. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1336–1340. doi: 10.1073/pnas.68.6.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Green M., Cartas M. The genome of RNA tumor viruses contains polyadenylic acid sequences. Proc Natl Acad Sci U S A. 1972 Apr;69(4):791–794. doi: 10.1073/pnas.69.4.791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Lim L., Canellakis E. S. Adenine-rich polymer associated with rabbit reticulocyte messenger RNA. Nature. 1970 Aug 15;227(5259):710–712. doi: 10.1038/227710a0. [DOI] [PubMed] [Google Scholar]
  14. Lim L., Canellakis Z. N., Canellakis E. S. Metabolism of naturally occurring homopolymers. II. Characterization of adenine-rich polynucleotides of mouse liver ribosomes. Biochim Biophys Acta. 1970 May 21;209(1):128–138. [PubMed] [Google Scholar]
  15. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  16. McGuire P. M., Swart C., Hodge L. D. Adenovirus messenger RNA in mammalian cells: failure of polyribosome association in the absence of nuclear cleavage. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1578–1582. doi: 10.1073/pnas.69.6.1578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mendecki J., Lee S. Y., Brawerman G. Characteristics of the polyadenylic acid segment associated with messenger ribonucleic acid in mouse sarcoma 180 ascites cells. Biochemistry. 1972 Feb 29;11(5):792–798. doi: 10.1021/bi00755a018. [DOI] [PubMed] [Google Scholar]
  18. Molloy G. R., Sporn M. B., Kelley D. E., Perry R. P. Localization of polyadenylic acid sequences in messenger ribonucleic acid of mammalian cells. Biochemistry. 1972 Aug 15;11(17):3256–3260. doi: 10.1021/bi00767a020. [DOI] [PubMed] [Google Scholar]
  19. Nakazato H., Edmonds M. The isolation and purification of rapidly labeled polysome-bound ribonucleic acid on polythymidylate cellulose. J Biol Chem. 1972 May 25;247(10):3365–3367. [PubMed] [Google Scholar]
  20. Pemberton R. E., Baglioni C. Duck hemoglobin messenger RNA contains a polynucleotide sequence rich in adenylic acid. J Mol Biol. 1972 Apr 14;65(3):531–535. doi: 10.1016/0022-2836(72)90207-0. [DOI] [PubMed] [Google Scholar]
  21. Perry R. P., La Torre J., Kelley D. E., Greenberg J. R. On the lability of poly(A) sequences during extraction of messenger RNA from polyribosomes. Biochim Biophys Acta. 1972 Mar 14;262(2):220–226. doi: 10.1016/0005-2787(72)90236-5. [DOI] [PubMed] [Google Scholar]
  22. Raff R. A., Colot H. V., Selvig S. E., Gross P. R. Oogenetic origin of messenger RNA for embryonic synthesis of microtubule proteins. Nature. 1972 Jan 28;235(5335):211–214. doi: 10.1038/235211a0. [DOI] [PubMed] [Google Scholar]
  23. Rosenfeld G. C., Comstock J. P., Means A. R., O'Malley B. W. A rapid method for the isolation and partial purification of specific eucaryotic messenger RNA's. Biochem Biophys Res Commun. 1972 Apr 28;47(2):387–392. doi: 10.1016/0006-291x(72)90725-5. [DOI] [PubMed] [Google Scholar]
  24. Rosenfeld M. G., Abrass J. B., Mendelsohn J., Ross B. A., Boone R. F., Garren L. D. Control of transcription of RNA rich in polyadenylic acid in human lymphocytes. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2306–2311. doi: 10.1073/pnas.69.8.2306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Selvig S. E., Gross P. R., Hunter A. L. Cytoplasmic synthesis of RNA in the sea urchin embryo. Dev Biol. 1970 Jun;22(2):343–365. doi: 10.1016/0012-1606(70)90158-2. [DOI] [PubMed] [Google Scholar]
  26. Sheldon R., Jurale C., Kates J. Detection of polyadenylic acid sequences in viral and eukaryotic RNA(polu(U)-cellulose columns-poly(U) filters-fiberglass-HeLa cells-bacteriophage T4). Proc Natl Acad Sci U S A. 1972 Feb;69(2):417–421. doi: 10.1073/pnas.69.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sheldon R., Kates J., Kelley D. E., Perry R. P. Polyadenylic acid sequences on 3' termini of vaccinia messenger ribonucleic acid and mammalian nuclear and messenger ribonucleic acid. Biochemistry. 1972 Sep 26;11(20):3829–3834. doi: 10.1021/bi00770a023. [DOI] [PubMed] [Google Scholar]
  28. Slater D. W., Slater I., Gillespie D. Post-fertilization synthesis of polyadenylic acid in sea urchin embryos. Nature. 1972 Dec 8;240(5380):333–337. doi: 10.1038/240333a0. [DOI] [PubMed] [Google Scholar]
  29. Slater D. W., Spiegelman S. A chemical and physical characterization of echinoid RNA during early embryogenesis. Biophys J. 2008 Dec 31;6(4):385–404. doi: 10.1016/S0006-3495(66)86665-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Slater D. W., Spiegelman S. An estimation of genetic messages in the unfertilized echinoid egg. Proc Natl Acad Sci U S A. 1966 Jul;56(1):164–170. doi: 10.1073/pnas.56.1.164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Slater D. W., Spiegelman S. Template capabilities and size distribution of echinoid RNA during early development. Biochim Biophys Acta. 1968 Aug 23;166(1):82–93. doi: 10.1016/0005-2787(68)90493-0. [DOI] [PubMed] [Google Scholar]
  32. Slater D. W., Spiegelman S. Transcriptive expression during sea urchin embryogenesis. Biochim Biophys Acta. 1970 Jul 16;213(1):194–207. doi: 10.1016/0005-2787(70)90020-1. [DOI] [PubMed] [Google Scholar]
  33. Slater I., Slater D. W. DNA polymerase potentials of sea urchin embryos. Nat New Biol. 1972 May 17;237(72):81–85. doi: 10.1038/newbio237081a0. [DOI] [PubMed] [Google Scholar]
  34. Soeiro R., Darnell J. E. Competition hybridization by "pre-saturation" of HeLa cell DNA. J Mol Biol. 1969 Sep 28;44(3):551–562. doi: 10.1016/0022-2836(69)90379-9. [DOI] [PubMed] [Google Scholar]
  35. Stevens R. H., Williamson A. R. Specific IgG mRNA molecules from myeloma cells in heterogeneous nuclear and cytoplasmic RNA containing poly-A. Nature. 1972 Sep 15;239(5368):143–146. doi: 10.1038/239143a0. [DOI] [PubMed] [Google Scholar]
  36. Swan D., Aviv H., Leder P. Purification and properties of biologically active messenger RNA for a myeloma light chain. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1967–1971. doi: 10.1073/pnas.69.7.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Terman S. A. Relative effect of transcription-level and translation-level control of protein synthesis during early development of the sea urchin. Proc Natl Acad Sci U S A. 1970 Apr;65(4):985–992. doi: 10.1073/pnas.65.4.985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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