Skip to main content
PMC home page
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
. 1988 Jul;85(13):4705–4709. doi: 10.1073/pnas.85.13.4705

Nuclear posttranscriptional processing of thymidine kinase mRNA at the onset of DNA synthesis.

J M Gudas 1, G B Knight 1, A B Pardee 1
PMCID: PMC280504  PMID: 2898781

Abstract

The posttranscriptional regulatory mechanism(s) underlying thymidine kinase (TK) mRNA accumulation was investigated in BALB/c 3T3 cells during their progression from G0 into S phase of the cell cycle. Very little TK mRNA could be detected in either the nuclear or the cytoplasmic compartment from cells harvested in G0 or G1. At the onset of S phase, however, the level of nuclear TK mRNA precursors and mature TK mRNAs increased dramatically. The high molecular weight TK heterogeneous nuclear RNA species detected in the nuclei of S-phase cells were polyadenylylated and hybridized to intron sequences derived from the TK gene. A series of high molecular weight precursors could be chased to lower molecular weight species in the presence of actinomycin D, suggesting an ordered removal of intron sequences with the kinetics of a precursor-product relationship. These results demonstrate a striking change in the nuclear posttranscriptional processing of TK heterogeneous nuclear RNA at the G1-S boundary and, furthermore, define a model system for the examination of RNA-processing events in vivo.

Full text

PDF
4705

Images in this article

4706Image
on p.4706
4706Image
on p.4706
4707Image
on p.4707
4707Image
on p.4707
4707Image
on p.4707
4707Image
on p.4707
4708Image
on p.4708

Selected References

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

  1. Anderson C. W., Lewis J. B., Atkins J. F., Gesteland R. F. Cell-free synthesis of adenovirus 2 proteins programmed by fractionated messenger RNA: a comparison of polypeptide products and messenger RNA lengths. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2756–2760. doi: 10.1073/pnas.71.7.2756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Artishevsky A., Wooden S., Sharma A., Resendez E., Jr, Lee A. S. Cell-cycle regulatory sequences in a hamster histone promoter and their interactions with cellular factors. 1987 Aug 27-Sep 2Nature. 328(6133):823–827. doi: 10.1038/328823a0. [DOI] [PubMed] [Google Scholar]
  3. Bradshaw H. D., Jr, Deininger P. L. Human thymidine kinase gene: molecular cloning and nucleotide sequence of a cDNA expressible in mammalian cells. Mol Cell Biol. 1984 Nov;4(11):2316–2320. doi: 10.1128/mcb.4.11.2316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradshaw H. D., Jr Molecular cloning and cell cycle-specific regulation of a functional human thymidine kinase gene. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5588–5591. doi: 10.1073/pnas.80.18.5588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caffarelli E., Fragapane P., Gehring C., Bozzoni I. The accumulation of mature RNA for the Xenopus laevis ribosomal protein L1 is controlled at the level of splicing and turnover of the precursor RNA. EMBO J. 1987 Nov;6(11):3493–3498. doi: 10.1002/j.1460-2075.1987.tb02674.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Campisi J., Gray H. E., Pardee A. B., Dean M., Sonenshein G. E. Cell-cycle control of c-myc but not c-ras expression is lost following chemical transformation. Cell. 1984 Feb;36(2):241–247. doi: 10.1016/0092-8674(84)90217-4. [DOI] [PubMed] [Google Scholar]
  7. Campisi J., Medrano E. E., Morreo G., Pardee A. B. Restriction point control of cell growth by a labile protein: evidence for increased stability in transformed cells. Proc Natl Acad Sci U S A. 1982 Jan;79(2):436–440. doi: 10.1073/pnas.79.2.436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Campisi J., Pardee A. B. An artifact in measurement of S phase initiation and its implication for the kinetics of S phase-specific enzyme activities. Exp Cell Res. 1982 Aug;140(2):389–393. doi: 10.1016/0014-4827(82)90128-8. [DOI] [PubMed] [Google Scholar]
  9. Campisi J., Pardee A. B. Post-transcriptional control of the onset of DNA synthesis by an insulin-like growth factor. Mol Cell Biol. 1984 Sep;4(9):1807–1814. doi: 10.1128/mcb.4.9.1807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Capasso O., Heintz N. Regulated expression of mammalian histone H4 genes in vivo requires a trans-acting transcription factor. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5622–5626. doi: 10.1073/pnas.82.17.5622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  12. Coppock D. L., Pardee A. B. Control of thymidine kinase mRNA during the cell cycle. Mol Cell Biol. 1987 Aug;7(8):2925–2932. doi: 10.1128/mcb.7.8.2925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Coppock D. L., Pardee A. B. Regulation of thymidine kinase activity in the cell cycle by a labile protein. J Cell Physiol. 1985 Aug;124(2):269–274. doi: 10.1002/jcp.1041240215. [DOI] [PubMed] [Google Scholar]
  14. Farnham P. J., Schimke R. T. Transcriptional regulation of mouse dihydrofolate reductase in the cell cycle. J Biol Chem. 1985 Jun 25;260(12):7675–7680. [PubMed] [Google Scholar]
  15. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  16. Flemington E., Bradshaw H. D., Jr, Traina-Dorge V., Slagel V., Deininger P. L. Sequence, structure and promoter characterization of the human thymidine kinase gene. Gene. 1987;52(2-3):267–277. doi: 10.1016/0378-1119(87)90053-9. [DOI] [PubMed] [Google Scholar]
  17. Goustin A. S., Leof E. B., Shipley G. D., Moses H. L. Growth factors and cancer. Cancer Res. 1986 Mar;46(3):1015–1029. [PubMed] [Google Scholar]
  18. Greenberg M. E., Ziff E. B. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene. Nature. 1984 Oct 4;311(5985):433–438. doi: 10.1038/311433a0. [DOI] [PubMed] [Google Scholar]
  19. Groudine M., Casimir C. Post-transcriptional regulation of the chicken thymidine kinase gene. Nucleic Acids Res. 1984 Feb 10;12(3):1427–1446. doi: 10.1093/nar/12.3.1427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hatzoglou M., Sekeris C. E., Hanson R. W. Processing of phosphoenolpyruvate carboxykinase (GTP) RNA in vivo. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4346–4350. doi: 10.1073/pnas.82.13.4346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hofbauer R., Müllner E., Seiser C., Wintersberger E. Cell cycle regulated synthesis of stable mouse thymidine kinase mRNA is mediated by a sequence within the cDNA. Nucleic Acids Res. 1987 Jan 26;15(2):741–752. doi: 10.1093/nar/15.2.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jenh C. H., Geyer P. K., Johnson L. F. Control of thymidylate synthase mRNA content and gene transcription in an overproducing mouse cell line. Mol Cell Biol. 1985 Oct;5(10):2527–2532. doi: 10.1128/mcb.5.10.2527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kelly K., Cochran B. H., Stiles C. D., Leder P. Cell-specific regulation of the c-myc gene by lymphocyte mitogens and platelet-derived growth factor. Cell. 1983 Dec;35(3 Pt 2):603–610. doi: 10.1016/0092-8674(83)90092-2. [DOI] [PubMed] [Google Scholar]
  24. Kit S. Thymidine kinase, DNA synthesis and cancer. Mol Cell Biochem. 1976 Jun 15;11(3):161–182. doi: 10.1007/BF01744997. [DOI] [PubMed] [Google Scholar]
  25. Knight G. B., Gudas J. M., Pardee A. B. Cell-cycle-specific interaction of nuclear DNA-binding proteins with a CCAAT sequence from the human thymidine kinase gene. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8350–8354. doi: 10.1073/pnas.84.23.8350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lewis J. A. Structure and expression of the Chinese hamster thymidine kinase gene. Mol Cell Biol. 1986 Jun;6(6):1998–2010. doi: 10.1128/mcb.6.6.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Leys E. J., Crouse G. F., Kellems R. E. Dihydrofolate reductase gene expression in cultured mouse cells is regulated by transcript stabilization in the nucleus. J Cell Biol. 1984 Jul;99(1 Pt 1):180–187. doi: 10.1083/jcb.99.1.180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lin P. F., Lieberman H. B., Yeh D. B., Xu T., Zhao S. Y., Ruddle F. H. Molecular cloning and structural analysis of murine thymidine kinase genomic and cDNA sequences. Mol Cell Biol. 1985 Nov;5(11):3149–3156. doi: 10.1128/mcb.5.11.3149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lin P. F., Zhao S. Y., Ruddle F. H. Genomic cloning and preliminary characterization of the human thymidine kinase gene. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6528–6532. doi: 10.1073/pnas.80.21.6528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Merrill G. F., Harland R. M., Groudine M., McKnight S. L. Genetic and physical analysis of the chicken tk gene. Mol Cell Biol. 1984 Sep;4(9):1769–1776. doi: 10.1128/mcb.4.9.1769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Murphy A. J., Efstratiadis A. Cloning vectors for expression of cDNA libraries in mammalian cells. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8277–8281. doi: 10.1073/pnas.84.23.8277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Müller R., Bravo R., Burckhardt J., Curran T. Induction of c-fos gene and protein by growth factors precedes activation of c-myc. Nature. 1984 Dec 20;312(5996):716–720. doi: 10.1038/312716a0. [DOI] [PubMed] [Google Scholar]
  33. Navalgund L. G., Rossana C., Muench A. J., Johnson L. F. Cell cycle regulation of thymidylate synthetase gene expression in cultured mouse fibroblasts. J Biol Chem. 1980 Aug 10;255(15):7386–7390. [PubMed] [Google Scholar]
  34. Pardee A. B. Molecules involved in proliferation of normal and cancer cells: presidential address. Cancer Res. 1987 Mar 15;47(6):1488–1491. [PubMed] [Google Scholar]
  35. Parnes J. R., Seidman J. G. Structure of wild-type and mutant mouse beta 2-microglobulin genes. Cell. 1982 Jun;29(2):661–669. doi: 10.1016/0092-8674(82)90182-9. [DOI] [PubMed] [Google Scholar]
  36. Parnes J. R., Sizer K. C., Seidman J. G., Stallings V., Hyman R. A mutational hot-spot within an intron of the mouse beta 2-microglobulin gene. EMBO J. 1986 Jan;5(1):103–111. doi: 10.1002/j.1460-2075.1986.tb04183.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Perucho M., Hanahan D., Lipsich L., Wigler M. Isolation of the chicken thymidine kinase gene by plasmid rescue. Nature. 1980 May 22;285(5762):207–210. doi: 10.1038/285207a0. [DOI] [PubMed] [Google Scholar]
  38. Plumb M., Stein J., Stein G. Coordinate regulation of multiple histone mRNAs during the cell cycle in HeLa cells. Nucleic Acids Res. 1983 Apr 25;11(8):2391–2410. doi: 10.1093/nar/11.8.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rossow P. W., Riddle V. G., Pardee A. B. Synthesis of labile, serum-dependent protein in early G1 controls animal cell growth. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4446–4450. doi: 10.1073/pnas.76.9.4446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Santiago C., Collins M., Johnson L. F. In vitro and in vivo analysis of the control of dihydrofolate reductase gene transcription in serum-stimulated mouse fibroblasts. J Cell Physiol. 1984 Jan;118(1):79–86. doi: 10.1002/jcp.1041180114. [DOI] [PubMed] [Google Scholar]
  41. Seiler-Tuyns A., Paterson B. M. Cell cycle regulation of a mouse histone H4 gene requires the H4 promoter. Mol Cell Biol. 1987 Mar;7(3):1048–1054. doi: 10.1128/mcb.7.3.1048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Shiels B. R., Northemann W., Gehring M. R., Fey G. H. Modified nuclear processing of alpha 1-acid glycoprotein RNA during inflammation. J Biol Chem. 1987 Sep 15;262(26):12826–12831. [PubMed] [Google Scholar]
  43. Stewart C. J., Ito M., Conrad S. E. Evidence for transcriptional and post-transcriptional control of the cellular thymidine kinase gene. Mol Cell Biol. 1987 Mar;7(3):1156–1163. doi: 10.1128/mcb.7.3.1156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Stuart P., Ito M., Stewart C., Conrad S. E. Induction of cellular thymidine kinase occurs at the mRNA level. Mol Cell Biol. 1985 Jun;5(6):1490–1497. doi: 10.1128/mcb.5.6.1490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tsai M. J., Ting A. C., Nordstrom J. L., Zimmer W., O'Malley B. W. Processing of high molecular weight ovalbumin and ovomucoid precursor RNAs to messenger RNA. Cell. 1980 Nov;22(1 Pt 1):219–230. doi: 10.1016/0092-8674(80)90170-1. [DOI] [PubMed] [Google Scholar]
  46. Vaessen R. T., Houweling A., van der Eb A. J. Post-transcriptional control of class I MHC mRNA expression in adenovirus 12-transformed cells. Science. 1987 Mar 20;235(4795):1486–1488. doi: 10.1126/science.3823900. [DOI] [PubMed] [Google Scholar]
  47. Wiedemann L. M., Johnson L. F. Regulation of dihydrofolate reductase synthesis in an overproducing 3T6 cell line during transition from resting to growing state. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2818–2822. doi: 10.1073/pnas.76.6.2818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wilkinson M. F., MacLeod C. L. Induction of T-cell receptor-alpha and -beta mRNA in SL12 cells can occur by transcriptional and post-transcriptional mechanisms. EMBO J. 1988 Jan;7(1):101–109. doi: 10.1002/j.1460-2075.1988.tb02788.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yang H. C., Pardee A. B. Insulin-like growth factor I regulation of transcription and replicating enzyme induction necessary for DNA synthesis. J Cell Physiol. 1986 Jun;127(3):410–416. doi: 10.1002/jcp.1041270309. [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