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. 1984 Dec 1;3(12):2787–2792. doi: 10.1002/j.1460-2075.1984.tb02210.x

Human globin gene transcription in injected Xenopus oocytes: enhancement by sodium butyrate.

G A Partington, N J Yarwood, T R Rutherford
PMCID: PMC557767  PMID: 6098445

Abstract

We have studied the expression of the human adult (beta) and foetal (gamma) globin genes by injecting a cosmid containing the complete -G gamma-A gamma-psi beta-delta-beta cluster into Xenopus oocytes. The transcripts from the gamma and beta genes were characterised with respect to their 5' termini by S1 nuclease mapping using probes which extend 5' to the cap site and 3' into the first exon. The only beta transcripts detectable in injected oocytes spanned from either -231 or -177 (cap site +1) into the first exon. The sensitivity of this transcription to inhibition by a low concentration of alpha-amanitin indicated that polymerase II was responsible. S1 analysis of gamma gene transcripts showed that the predominant transcript mapped from a site at -227, but a low level of transcription from the cap site was also detectable. Incubation of oocytes in saline solution containing sodium butyrate prior to injection resulted in a significant stimulation of transcription from the gamma gene cap site. In contrast, transcription from the beta gene upstream sites was only slightly increased. The possible role of sodium butyrate in promoting this effect is discussed in the context of its known property as a modifier of cellular chromatin.

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Selected References

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

  1. Adamson E. D., Woodland H. R. Histone synthesis in early amphibian development: histone and DNA syntheses are not co-ordinated. J Mol Biol. 1974 Sep 15;88(2):263–285. doi: 10.1016/0022-2836(74)90481-1. [DOI] [PubMed] [Google Scholar]
  2. Allan M., Lanyon W. G., Paul J. Multiple origins of transcription in the 4.5 Kb upstream of the epsilon-globin gene. Cell. 1983 Nov;35(1):187–197. doi: 10.1016/0092-8674(83)90221-0. [DOI] [PubMed] [Google Scholar]
  3. Andersson L. C., Jokinen M., Gahmberg C. G. Induction of erythroid differentiation in the human leukaemia cell line K562. Nature. 1979 Mar 22;278(5702):364–365. doi: 10.1038/278364a0. [DOI] [PubMed] [Google Scholar]
  4. Asselbergs F. A., Smart J. E., Mathews M. B. Analysis of expression of adenovirus DNA (fragments) by microinjection in Xenopus oocytes. Independent synthesis of minor early region 2 proteins. J Mol Biol. 1983 Jan 15;163(2):209–238. doi: 10.1016/0022-2836(83)90004-9. [DOI] [PubMed] [Google Scholar]
  5. Bentley D. L., Farrell P. J., Rabbitts T. H. Unrearranged immunoglobulin variable region genes have a functional promoter. Nucleic Acids Res. 1982 Mar 25;10(6):1841–1856. doi: 10.1093/nar/10.6.1841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Benyajati C., Worcel A. Isolation, characterization, and structure of the folded interphase genome of Drosophila melanogaster. Cell. 1976 Nov;9(3):393–407. doi: 10.1016/0092-8674(76)90084-2. [DOI] [PubMed] [Google Scholar]
  7. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  8. Bode J., Gómez-Lira M. M., Schröter H. Nucleosomal particles open as the histone core becomes hyperacetylated. Eur J Biochem. 1983 Feb 15;130(3):437–445. doi: 10.1111/j.1432-1033.1983.tb07170.x. [DOI] [PubMed] [Google Scholar]
  9. Candido E. P., Reeves R., Davie J. R. Sodium butyrate inhibits histone deacetylation in cultured cells. Cell. 1978 May;14(1):105–113. doi: 10.1016/0092-8674(78)90305-7. [DOI] [PubMed] [Google Scholar]
  10. Carlson D. P., Ross J. Human beta-globin promoter and coding sequences transcribed by RNA polymerase III. Cell. 1983 Oct;34(3):857–864. doi: 10.1016/0092-8674(83)90543-3. [DOI] [PubMed] [Google Scholar]
  11. D'Anna J. A., Tobey R. A., Gurley L. R. Concentration-dependent effects of sodium butyrate in Chinese hamster cells: cell-cycle progression, inner-histone acetylation, histone H1 dephosphorylation, and induction of an H1-like protein. Biochemistry. 1980 Jun 10;19(12):2656–2671. doi: 10.1021/bi00553a019. [DOI] [PubMed] [Google Scholar]
  12. Davie J. R., Saunders C. A. Chemical composition of nucleosomes among domains of calf thymus chromatin differing in micrococcal nuclease accessibility and solubility properties. J Biol Chem. 1981 Dec 10;256(23):12574–12580. [PubMed] [Google Scholar]
  13. De Robertis E. M., Olson M. V. Transcription and processing of cloned yeast tyrosine tRNA genes microinjected into frog oocytes. Nature. 1979 Mar 8;278(5700):137–143. doi: 10.1038/278137a0. [DOI] [PubMed] [Google Scholar]
  14. Dierks P., van Ooyen A., Cochran M. D., Dobkin C., Reiser J., Weissmann C. Three regions upstream from the cap site are required for efficient and accurate transcription of the rabbit beta-globin gene in mouse 3T6 cells. Cell. 1983 Mar;32(3):695–706. doi: 10.1016/0092-8674(83)90055-7. [DOI] [PubMed] [Google Scholar]
  15. Dumont J. N. Oogenesis in Xenopus laevis (Daudin). I. Stages of oocyte development in laboratory maintained animals. J Morphol. 1972 Feb;136(2):153–179. doi: 10.1002/jmor.1051360203. [DOI] [PubMed] [Google Scholar]
  16. Egan P. A., Levy-Wilson B. Structure of transcriptionally active and inactive nucleosomes from butyrate-treated and control HeLa cells. Biochemistry. 1981 Jun 23;20(13):3695–3702. doi: 10.1021/bi00516a005. [DOI] [PubMed] [Google Scholar]
  17. Elgin S. C. DNAase I-hypersensitive sites of chromatin. Cell. 1981 Dec;27(3 Pt 2):413–415. doi: 10.1016/0092-8674(81)90381-0. [DOI] [PubMed] [Google Scholar]
  18. Emerson B. M., Felsenfeld G. Specific factor conferring nuclease hypersensitivity at the 5' end of the chicken adult beta-globin gene. Proc Natl Acad Sci U S A. 1984 Jan;81(1):95–99. doi: 10.1073/pnas.81.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gargiulo G., Worcel A. Analysis of the chromatin assembled in germinal vesicles of Xenopus oocytes. J Mol Biol. 1983 Nov 5;170(3):699–722. doi: 10.1016/s0022-2836(83)80128-4. [DOI] [PubMed] [Google Scholar]
  20. Gorman C. M., Howard B. H., Reeves R. Expression of recombinant plasmids in mammalian cells is enhanced by sodium butyrate. Nucleic Acids Res. 1983 Nov 11;11(21):7631–7648. doi: 10.1093/nar/11.21.7631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Grindlay G. J., Lanyon W. G., Allan M., Paul J. Alternative sites of transcription initiation upstream of the canonical cap site in human gamma-globin and beta-globin genes. Nucleic Acids Res. 1984 Feb 24;12(4):1811–1820. doi: 10.1093/nar/12.4.1811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Grosveld F. G., Lund T., Murray E. J., Mellor A. L., Dahl H. H., Flavell R. A. The construction of cosmid libraries which can be used to transform eukaryotic cells. Nucleic Acids Res. 1982 Nov 11;10(21):6715–6732. doi: 10.1093/nar/10.21.6715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Grosveld G. C., de Boer E., Shewmaker C. K., Flavell R. A. DNA sequences necessary for transcription of the rabbit beta-globin gene in vivo. Nature. 1982 Jan 14;295(5845):120–126. doi: 10.1038/295120a0. [DOI] [PubMed] [Google Scholar]
  24. Groudine M., Kohwi-Shigematsu T., Gelinas R., Stamatoyannopoulos G., Papayannopoulou T. Human fetal to adult hemoglobin switching: changes in chromatin structure of the beta-globin gene locus. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7551–7555. doi: 10.1073/pnas.80.24.7551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Gurdon J. B., Melton D. A. Gene transfer in amphibian eggs and oocytes. Annu Rev Genet. 1981;15:189–218. doi: 10.1146/annurev.ge.15.120181.001201. [DOI] [PubMed] [Google Scholar]
  26. Harland R. M., Weintraub H., McKnight S. L. Transcription of DNA injected into Xenopus oocytes is influenced by template topology. Nature. 1983 Mar 3;302(5903):38–43. doi: 10.1038/302038a0. [DOI] [PubMed] [Google Scholar]
  27. Humphries R. K., Ley T., Turner P., Moulton A. D., Nienhuis A. W. Differences in human alpha-, beta- and delta-globin gene expression in monkey kidney cells. Cell. 1982 Aug;30(1):173–183. doi: 10.1016/0092-8674(82)90023-x. [DOI] [PubMed] [Google Scholar]
  28. Korn L. J. Transcription of Xenopus 5S ribosomal RNA genes. Nature. 1982 Jan 14;295(5845):101–105. doi: 10.1038/295101a0. [DOI] [PubMed] [Google Scholar]
  29. Lachman H. M., Mears J. G. DNase I hypersensitivity in the gamma globin gene locus of K562 cells. Nucleic Acids Res. 1983 Sep 10;11(17):6065–6077. doi: 10.1093/nar/11.17.6065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Leder A., Leder P. Butyric acid, a potent inducer of erythroid differentiation in cultured erythroleukemic cells. Cell. 1975 Jul;5(3):319–322. doi: 10.1016/0092-8674(75)90107-5. [DOI] [PubMed] [Google Scholar]
  31. Levy-Wilson B. Enhanced phosphorylation of high-mobility-group proteins in nuclease-sensitive mononucleosomes from butyrate-treated HeLa cells. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2189–2193. doi: 10.1073/pnas.78.4.2189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ley T. J., Nienhuis A. W. A weak upstream promoter gives rise to long human beta-globin RNA molecules. Biochem Biophys Res Commun. 1983 May 16;112(3):1041–1048. doi: 10.1016/0006-291x(83)91723-0. [DOI] [PubMed] [Google Scholar]
  33. McKnight S. L., Kingsbury R. Transcriptional control signals of a eukaryotic protein-coding gene. Science. 1982 Jul 23;217(4557):316–324. doi: 10.1126/science.6283634. [DOI] [PubMed] [Google Scholar]
  34. McKnight S. L. The nucleotide sequence and transcript map of the herpes simplex virus thymidine kinase gene. Nucleic Acids Res. 1980 Dec 20;8(24):5949–5964. doi: 10.1093/nar/8.24.5949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Mertz J. E., Gurdon J. B. Purified DNAs are transcribed after microinjection into Xenopus oocytes. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1502–1506. doi: 10.1073/pnas.74.4.1502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Mertz J. E. Linear DNA does not form chromatin containing regularly spaced nucleosomes. Mol Cell Biol. 1982 Dec;2(12):1608–1618. doi: 10.1128/mcb.2.12.1608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Miller T. J., Mertz J. E. Template structural requirements for transcription in vivo by RNA polymerase II. Mol Cell Biol. 1982 Dec;2(12):1595–1607. doi: 10.1128/mcb.2.12.1595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Miller T. J., Stephens D. L., Mertz J. E. Kinetics of accumulation and processing of simian virus 40 RNA in Xenopus laevis oocytes injected with simian virus 40 DNA. Mol Cell Biol. 1982 Dec;2(12):1581–1594. doi: 10.1128/mcb.2.12.1581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Osborne T. F., Berk A. J. Far upstream initiation sites for adenovirus early region 1A transcription are utilized after the onset of viral DNA replication. J Virol. 1983 Feb;45(2):594–599. doi: 10.1128/jvi.45.2.594-599.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Perry M., Chalkley R. Histone acetylation increases the solubility of chromatin and occurs sequentially over most of the chromatin. A novel model for the biological role of histone acetylation. J Biol Chem. 1982 Jul 10;257(13):7336–7347. [PubMed] [Google Scholar]
  41. Pogo B. G., Pogo A. O., Allfrey V. G., Mirsky A. E. Changing patterns of histone acetylation and RNA synthesis in regeneration of the liver. Proc Natl Acad Sci U S A. 1968 Apr;59(4):1337–1344. doi: 10.1073/pnas.59.4.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Poncz M., Schwartz E., Ballantine M., Surrey S. Nucleotide sequence analysis of the delta beta-globin gene region in humans. J Biol Chem. 1983 Oct 10;258(19):11599–11609. [PubMed] [Google Scholar]
  43. Probst E., Kressmann A., Birnstiel M. L. Expression of sea urchin histone genes in the oocyte of Xenopus laevis. J Mol Biol. 1979 Dec 15;135(3):709–732. doi: 10.1016/0022-2836(79)90173-6. [DOI] [PubMed] [Google Scholar]
  44. Proudfoot N. J., Rutherford T. R., Partington G. A. Transcriptional analysis of human zeta globin genes. EMBO J. 1984 Jul;3(7):1533–1540. doi: 10.1002/j.1460-2075.1984.tb02007.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sassone-Corsi P., Hen R., Borrelli E., Leff T., Chambon P. Far upstream sequences are required for efficient transcription from the adenovirus-2 E1A transcription unit. Nucleic Acids Res. 1983 Dec 20;11(24):8735–8745. doi: 10.1093/nar/11.24.8735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sealy L., Chalkley R. The effect of sodium butyrate on histone modification. Cell. 1978 May;14(1):115–121. doi: 10.1016/0092-8674(78)90306-9. [DOI] [PubMed] [Google Scholar]
  47. Shen S. H., Slightom J. L., Smithies O. A history of the human fetal globin gene duplication. Cell. 1981 Oct;26(2 Pt 2):191–203. doi: 10.1016/0092-8674(81)90302-0. [DOI] [PubMed] [Google Scholar]
  48. Simpson R. T. Structure of chromatin containing extensively acetylated H3 and H4. Cell. 1978 Apr;13(4):691–699. doi: 10.1016/0092-8674(78)90219-2. [DOI] [PubMed] [Google Scholar]
  49. Trendelenburg M. F., Mathis D., Oudet P. Transcription units of chicken ovalbumin gene observed after injection of cloned complete genes into Xenopus oocyte nuclei. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5984–5988. doi: 10.1073/pnas.77.10.5984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Vidali G., Boffa L. C., Bradbury E. M., Allfrey V. G. Butyrate suppression of histone deacetylation leads to accumulation of multiacetylated forms of histones H3 and H4 and increased DNase I sensitivity of the associated DNA sequences. Proc Natl Acad Sci U S A. 1978 May;75(5):2239–2243. doi: 10.1073/pnas.75.5.2239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Weisbrod S. Active chromatin. Nature. 1982 May 27;297(5864):289–295. doi: 10.1038/297289a0. [DOI] [PubMed] [Google Scholar]
  53. Wickens M. P., Gurdon J. B. Post-transcriptional processing of simian virus 40 late transcripts in injected frog oocytes. J Mol Biol. 1983 Jan 5;163(1):1–26. doi: 10.1016/0022-2836(83)90027-x. [DOI] [PubMed] [Google Scholar]
  54. Wickens M. P., Woo S., O'Malley B. W., Gurdon J. B. Expression of a chicken chromosomal ovalbumin gene injected into frog oocyte nuclei. Nature. 1980 Jun 26;285(5767):628–634. doi: 10.1038/285628a0. [DOI] [PubMed] [Google Scholar]
  55. Woodland H. R. The modification of stored histones H3 and H4 during the oogenesis and early development of Xenopus laevis. Dev Biol. 1979 Feb;68(2):360–370. doi: 10.1016/0012-1606(79)90210-0. [DOI] [PubMed] [Google Scholar]
  56. Wyllie A. H., Laskey R. A., Finch J., Gurdon J. B. Selective DNA conservation and chromatin assembly after injection of SV40 DNA into Xenopus oocytes. Dev Biol. 1978 May;64(1):178–188. doi: 10.1016/0012-1606(78)90069-6. [DOI] [PubMed] [Google Scholar]

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