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
. 1978 Nov;75(11):5344–5348. doi: 10.1073/pnas.75.11.5344

In vitro RNA-RNA splicing in adenovirus 2 mRNA formation.

J M Blanchard, J Weber, W Jelinek, J E Darnell
PMCID: PMC392959  PMID: 281684

Abstract

"Splicing" of the precursor to an adenovirus mRNA was accomplished in isolated cell-free extracts. Nuclei were prepared from hypotonically swollen cells that had been labeled with [3H]uridine for 10 min prior to nuclear isolation. Addition of a "cytoplasmic" fraction was required for the splicing to occur. The nuclear precursor, a poly(A)-terminated RNA molecule approximately 5 kilobases long, contained sequences complementary to the 58.5--75.9 region of the adenovirus 2 genome, including those sequences spliced out of the mature mRNA molecule. The in vitro spliced product was a poly(A)-terminated RNA molecule identical in size to the cytoplasmic 72,000 Mr protein mRNA (2 kilobases long) in which the sequences encoded in the 70.7--75.9 region of the viral genome were spliced to those encoded at 58.7--65.6, with the sequences encoded at 66.1--70.7 deleted.

Full text

PDF
5345

Selected References

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

  1. Aloni Y., Dhar R., Laub O., Horowitz M., Khoury G. Novel mechanism for RNA maturation: the leader sequences of simian virus 40 mRNA are not transcribed adjacent to the coding sequences. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3686–3690. doi: 10.1073/pnas.74.9.3686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersson J., Coutinho A., Lernhardt W., Melchers F. Clonal growth and maturation to immunoglobulin secretion in vitro of every growth-inducible B lymphocyte. Cell. 1977 Jan;10(1):27–34. doi: 10.1016/0092-8674(77)90136-2. [DOI] [PubMed] [Google Scholar]
  3. Bachenheimer S., Darnell J. E. Adenovirus-2 mRNA is transcribed as part of a high-molecular-weight precursor RNA. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4445–4449. doi: 10.1073/pnas.72.11.4445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bastos R. N., Aviv H. Globin RNA precursor molecules: biosynthesis and process in erythroid cells. Cell. 1977 Jul;11(3):641–650. doi: 10.1016/0092-8674(77)90081-2. [DOI] [PubMed] [Google Scholar]
  5. Berget S. M., Moore C., Sharp P. A. Spliced segments at the 5' terminus of adenovirus 2 late mRNA. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3171–3175. doi: 10.1073/pnas.74.8.3171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Berk A. J., Sharp P. A. Structure of the adenovirus 2 early mRNAs. Cell. 1978 Jul;14(3):695–711. doi: 10.1016/0092-8674(78)90252-0. [DOI] [PubMed] [Google Scholar]
  8. Berk A. J., Sharp P. A. Ultraviolet mapping of the adenovirus 2 early promoters. Cell. 1977 Sep;12(1):45–55. doi: 10.1016/0092-8674(77)90184-2. [DOI] [PubMed] [Google Scholar]
  9. Breathnach R., Mandel J. L., Chambon P. Ovalbumin gene is split in chicken DNA. Nature. 1977 Nov 24;270(5635):314–319. doi: 10.1038/270314a0. [DOI] [PubMed] [Google Scholar]
  10. Chow L. T., Gelinas R. E., Broker T. R., Roberts R. J. An amazing sequence arrangement at the 5' ends of adenovirus 2 messenger RNA. Cell. 1977 Sep;12(1):1–8. doi: 10.1016/0092-8674(77)90180-5. [DOI] [PubMed] [Google Scholar]
  11. Chow L. T., Roberts J. M., Lewis J. B., Broker T. R. A map of cytoplasmic RNA transcripts from lytic adenovirus type 2, determined by electron microscopy of RNA:DNA hybrids. Cell. 1977 Aug;11(4):819–836. doi: 10.1016/0092-8674(77)90294-x. [DOI] [PubMed] [Google Scholar]
  12. Cooke H. J., McKay R. D. Evolution of a human Y chromosome-specific repeated sequence. Cell. 1978 Mar;13(3):453–460. doi: 10.1016/0092-8674(78)90319-7. [DOI] [PubMed] [Google Scholar]
  13. Craig E. A., Raskas H. J. Nuclear transcripts larger than the cytoplasmic mRNAs are specified by segments of the adenovirus genome coding for early functions. Cell. 1976 Jun;8(2):205–213. doi: 10.1016/0092-8674(76)90004-0. [DOI] [PubMed] [Google Scholar]
  14. Curtis P. J., Mantei N., van den Berg J., Weissmann C. Presence of a putative 15S precursor to beta-globin mRNA but not to alpha-globin mRNA in Friend cells. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3184–3188. doi: 10.1073/pnas.74.8.3184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Eliceiri G. L., Gurney T., Jr Subcellular location of precursors to small nuclear RNA species C and D and of newly synthesized 5 S RNA in HeLa cells. Biochem Biophys Res Commun. 1978 Apr 14;81(3):915–919. doi: 10.1016/0006-291x(78)91438-9. [DOI] [PubMed] [Google Scholar]
  16. Evans R. M., Fraser N., Ziff E., Weber J., Wilson M., Darnell J. E. The initiation sites for RNA transcription in Ad2 DNA. Cell. 1977 Nov;12(3):733–739. doi: 10.1016/0092-8674(77)90273-2. [DOI] [PubMed] [Google Scholar]
  17. Flint J. The topography and transcription of the adenovirus genome. Cell. 1977 Feb;10(2):153–166. doi: 10.1016/0092-8674(77)90211-2. [DOI] [PubMed] [Google Scholar]
  18. Foster D. N., Gurney T., Jr Nuclear location of mammalian DNA polymerase activities. J Biol Chem. 1976 Dec 25;251(24):7893–7898. [PubMed] [Google Scholar]
  19. Gilmore-Hebert M., Wall R. Immunoglobulin light chain mRNA is processed from large nuclear RNA. Proc Natl Acad Sci U S A. 1978 Jan;75(1):342–345. doi: 10.1073/pnas.75.1.342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Goldberg S., Weber J., Darnell J. E., Jr The definition of a large viral transcription unit late in Ad2 infection of HeLa cells: mapping by effects of ultraviolet irradiation. Cell. 1977 Apr;10(4):617–621. doi: 10.1016/0092-8674(77)90094-0. [DOI] [PubMed] [Google Scholar]
  21. Goodman H. M., Olson M. V., Hall B. D. Nucleotide sequence of a mutant eukaryotic gene: the yeast tyrosine-inserting ochre suppressor SUP4-o. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5453–5457. doi: 10.1073/pnas.74.12.5453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hsu M. T., Ford J. Sequence arrangement of the 5' ends of simian virus 40 16S and 19S mRNAs. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4982–4985. doi: 10.1073/pnas.74.11.4982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jeffreys A. J., Flavell R. A. The rabbit beta-globin gene contains a large large insert in the coding sequence. Cell. 1977 Dec;12(4):1097–1108. doi: 10.1016/0092-8674(77)90172-6. [DOI] [PubMed] [Google Scholar]
  24. Jelinek W., Molloy G., Fernandez-Munoz R., Salditt M., Darnell J. E. Secondary structure in heterogeneous nuclear RNA: involvement of regions from repeated DNA sites. J Mol Biol. 1974 Jan 25;82(3):361–370. doi: 10.1016/0022-2836(74)90597-x. [DOI] [PubMed] [Google Scholar]
  25. Kitchingman G. R., Lai S. P., Westphal H. Loop structures in hybrids of early RNA and the separated strands of adenovirus DNA. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4392–4395. doi: 10.1073/pnas.74.10.4392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Klessig D. F. Two adenovirus mRNAs have a common 5' terminal leader sequence encoded at least 10 kb upstream from their main coding regions. Cell. 1977 Sep;12(1):9–21. doi: 10.1016/0092-8674(77)90181-7. [DOI] [PubMed] [Google Scholar]
  27. Knapp G., Beckmann J. S., Johnson P. F., Fuhrman S. A., Abelson J. Transcription and processing of intervening sequences in yeast tRNA genes. Cell. 1978 Jun;14(2):221–236. doi: 10.1016/0092-8674(78)90109-5. [DOI] [PubMed] [Google Scholar]
  28. Lewis J. B., Atkins J. F., Baum P. R., Solem R., Gesteland R. F., Anderson C. W. Location and identification of the genes for adenovirus type 2 early polypeptides. Cell. 1976 Jan;7(1):141–151. doi: 10.1016/0092-8674(76)90264-6. [DOI] [PubMed] [Google Scholar]
  29. Lönn U. Exclusive nuclear location of precursor 4 S RNA in vivo. J Mol Biol. 1977 Jun 5;112(4):661–666. doi: 10.1016/s0022-2836(77)80172-1. [DOI] [PubMed] [Google Scholar]
  30. Nevins J. R., Darnell J. E. Groups of adenovirus type 2 mRNA's derived from a large primary transcript: probable nuclear origin and possible common 3' ends. J Virol. 1978 Mar;25(3):811–823. doi: 10.1128/jvi.25.3.811-823.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Penman S., Scherrer K., Becker Y., Darnell J. E. POLYRIBOSOMES IN NORMAL AND POLIOVIRUS-INFECTED HELA CELLS AND THEIR RELATIONSHIP TO MESSENGER-RNA. Proc Natl Acad Sci U S A. 1963 May;49(5):654–662. doi: 10.1073/pnas.49.5.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Tilghman S. M., Tiemeier D. C., Seidman J. G., Peterlin B. M., Sullivan M., Maizel J. V., Leder P. Intervening sequence of DNA identified in the structural portion of a mouse beta-globin gene. Proc Natl Acad Sci U S A. 1978 Feb;75(2):725–729. doi: 10.1073/pnas.75.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tonegawa S., Maxam A. M., Tizard R., Bernard O., Gilbert W. Sequence of a mouse germ-line gene for a variable region of an immunoglobulin light chain. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1485–1489. doi: 10.1073/pnas.75.3.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Valenzuela P., Venegas A., Weinberg F., Bishop R., Rutter W. J. Structure of yeast phenylalanine-tRNA genes: an intervening DNA segment within the region coding for the tRNA. Proc Natl Acad Sci U S A. 1978 Jan;75(1):190–194. doi: 10.1073/pnas.75.1.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wall R., Darnell J. E. Presence of cell and virus specific sequences in the same molecules of nuclear RNA from virus transformed cells. Nat New Biol. 1971 Jul 21;232(29):73–76. doi: 10.1038/newbio232073a0. [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