Skip to main content
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
. 1977 Jan;74(1):178–182. doi: 10.1073/pnas.74.1.178

Purification of a putative precursor of globin messenger RNA from mouse nucleated erythroid cells.

S P Kwan, T G Wood, J B Lingrel
PMCID: PMC393221  PMID: 264673

Abstract

Nucleated erythroid cells were incubated for 10 min in the presence of [5-3H]uridine, and the total RNA was isolated by three different extraction procedures. RNA containing globin messenger RNA sequences was purified from other cellular RNAs by selective hybridization to globin complementary DNA cellulose. Depending upon the extraction procedure employed, 0.4-0.6% of the radioactively-labeled total cellular RNA applied to the column annealed to globin complementary DNA cellulose. The annealed RNA was treated with formaldehyde and analyzed by formaldehyde/polyacrylamide gel electrophoresis. Mature globin mRNA and an RNA migrating at approximately 15 S were observed. No globin mRNA containing sequences larger than 20 S were present. The 15S RNA was partially resolved from mature globin mRNA by neutral sucrose density gradient centrifugation. The RNA isolated from the heavy region of this gradient migrated as 15 S in the formaldehyde/polyacrylamide gels and retained its ability to quantitatively anneal to globin complementary DNA cellulose. On the basis of these observations, we conclude that nucleated erythroid cells obtained from the spleens of anemic mice have a 15S RNA which contains globin mRNA sequences. The 15S RNA is not an aggregate and is a good candidate for a globin mRNA precursor.

Full text

PDF
178

Selected References

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

  1. Curtis P. J., Weissmann C. Purification of globin messenger RNA from dimethylsulfoxide-induced Friend cells and detection of a putative globin messenger RNA precursor. J Mol Biol. 1976 Oct 5;106(4):1067–1075. doi: 10.1016/0022-2836(76)90353-3. [DOI] [PubMed] [Google Scholar]
  2. Gorski J., Morrison M. R., Merkel C. G., Lingrel J. B. Size heterogeneity of polyadenylate sequences in mouse globin messenger RNA. J Mol Biol. 1974 Jun 25;86(2):363–371. doi: 10.1016/0022-2836(74)90025-4. [DOI] [PubMed] [Google Scholar]
  3. Holmes D. S., Bonner J. Preparation, molecular weight, base composition, and secondary structure of giant nuclear ribonucleic acid. Biochemistry. 1973 Jun 5;12(12):2330–2338. doi: 10.1021/bi00736a023. [DOI] [PubMed] [Google Scholar]
  4. Knöchel W., Tiedemann H. Size distribution and cell-free translation of globin-coding HnRNA from avian erythroblasts. Biochim Biophys Acta. 1975 Feb 10;378(3):383–393. doi: 10.1016/0005-2787(75)90183-5. [DOI] [PubMed] [Google Scholar]
  5. Lee S. Y., Krsmanovic V., Brawerman G. Attachment of ribosomes to membranes during polysome formation in mouse sarcoma 180 cells. J Cell Biol. 1971 Jun;49(3):683–691. doi: 10.1083/jcb.49.3.683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lengyel J., Penman S. hnRNA size and processing as related to different DNA content in two dipterans: Drosophila and Aedes. Cell. 1975 Jul;5(3):281–290. doi: 10.1016/0092-8674(75)90103-8. [DOI] [PubMed] [Google Scholar]
  7. Lewin B. Units of transcription and translation: sequence components of heterogeneous nuclear RNA and messenger RNA. Cell. 1975 Feb;4(2):77–93. doi: 10.1016/0092-8674(75)90113-0. [DOI] [PubMed] [Google Scholar]
  8. Lewin B. Units of transcription and translation: the relationship between heterogeneous nuclear RNA and messenger RNA. Cell. 1975 Jan;4(1):11–20. doi: 10.1016/0092-8674(75)90128-2. [DOI] [PubMed] [Google Scholar]
  9. Melli M., Pemberton R. E. New method of studying the precursor-product relationship between high molecular weight RNA and messenger RNA. Nat New Biol. 1972 Apr 12;236(67):172–174. doi: 10.1038/newbio236172a0. [DOI] [PubMed] [Google Scholar]
  10. Merkel C. G., Kwan S., Lingrel J. B. Size of the polyadenylic acid region of newly synthesized globin messenger ribonucleic acid. J Biol Chem. 1975 May 25;250(10):3725–3728. [PubMed] [Google Scholar]
  11. Ojala D., Attardi G. Identification and partial characterization of multiple discrete polyadenylic acid containing RNA components coded for by HeLa cell mitochondrial DNA. J Mol Biol. 1974 Sep 5;88(1):205–219. doi: 10.1016/0022-2836(74)90305-2. [DOI] [PubMed] [Google Scholar]
  12. Ostertag W., Gaedicke G., Kluge N., Melderis H., Weimann B., Dube S. K. Globin messenger in mouse leukemic cells: activity associated with RNA species in the region of 8 to 16 S. FEBS Lett. 1973 Jun 1;32(2):219–222. doi: 10.1016/0014-5793(73)80836-1. [DOI] [PubMed] [Google Scholar]
  13. Perry R. P., Kelley D. E. Methylated constituents of heterogeneous nuclear RNA: presence in blocked 5' terminal structures. Cell. 1975 Sep;6(1):13–19. doi: 10.1016/0092-8674(75)90068-9. [DOI] [PubMed] [Google Scholar]
  14. Ross J. A precursor of globin messenger RNA. J Mol Biol. 1976 Sep 15;106(2):403–420. doi: 10.1016/0022-2836(76)90093-0. [DOI] [PubMed] [Google Scholar]
  15. Ruiz-Carrilo A., Beato M., Schutz G., Feigelson P., Allfrey V. G. Cell-free translation of the globin message within polydisperse high-molecular-weight ribonucleic acid of avian erythrocytes. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3641–3645. doi: 10.1073/pnas.70.12.3641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Russell E. S., McFarland E. C. Genetics of mouse hemoglobins. Ann N Y Acad Sci. 1974 Nov 29;241(0):25–38. doi: 10.1111/j.1749-6632.1974.tb21864.x. [DOI] [PubMed] [Google Scholar]
  17. SCHERRER K., DARNELL J. E. Sedimentation characteristics of rapidly labelled RNA from HeLa cells. Biochem Biophys Res Commun. 1962 Jun 4;7:486–490. doi: 10.1016/0006-291x(62)90341-8. [DOI] [PubMed] [Google Scholar]
  18. Salditt-Georgieff M., Jelinek W., Darnell J. E., Furuichi Y., Morgan M., Shatkin A. Methyl labeling of HeLa cell hnRNA: a comparison with mRNA. Cell. 1976 Feb;7(2):227–237. doi: 10.1016/0092-8674(76)90022-2. [DOI] [PubMed] [Google Scholar]
  19. Suzuki Y., Brown D. D. Isolation and identification of the messenger RNA for silk fibroin from Bombyx mori. J Mol Biol. 1972 Feb 14;63(3):409–429. doi: 10.1016/0022-2836(72)90437-8. [DOI] [PubMed] [Google Scholar]
  20. Venetianer P., Leder P. Enzymatic synthesis of solid phase-bound DNA sequences corresponding to specific mammalian genes. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3892–3895. doi: 10.1073/pnas.71.10.3892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Williamson R., Drewienkiewicz C. E., Paul J. Globin messenger sequences in high molecular weight RNA from embryonic mouse liver. Nat New Biol. 1973 Jan 17;241(107):66–68. doi: 10.1038/newbio241066a0. [DOI] [PubMed] [Google Scholar]
  22. Zimmerman S. B., Sandeen D. The ribonuclease activity of crystallized pancreatic deoxyribonuclease. Anal Biochem. 1966 Feb;14(2):269–277. doi: 10.1016/0003-2697(66)90137-0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES