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. 1986 Dec 1;5(12):3185–3193. doi: 10.1002/j.1460-2075.1986.tb04628.x

Upstream sequences required for tissue-specific activation of the cardiac actin gene in Xenopus laevis embryos.

T J Mohun, N Garrett, J B Gurdon
PMCID: PMC1167311  PMID: 3816759

Abstract

The entire DNA sequence of the Xenopus laevis cardiac actin gene was determined. A recombinant plasmid comprising the cardiac actin gene promoter fused to the bacterial chloramphenicol acetyl transferase (CAT) gene is correctly regulated after introduction into fertilized Xenopus eggs. The fusion gene shows a temporal and tissue-specific pattern of expression in the early embryo which is indistinguishable from that of the endogenous cardiac actin gene. The fusion gene is also activated in cultured embryo fragments that are induced by cell interactions to form embryonic muscle tissue. Tissue-specific expression of the recombinant requires sequences between 217 and 416 nucleotides upstream from the transcription initiation site. In contrast, both the chimaeric gene and the entire cardiac actin gene are expressed at a basal level after microinjection into Xenopus oocytes, requiring only the presence of a TATA box upstream from the cap site.

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Selected References

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

  1. Andres A. C., Muellener D. B., Ryffel G. U. Persistence, methylation and expression of vitellogenin gene derivatives after injection into fertilized eggs of Xenopus laevis. Nucleic Acids Res. 1984 Mar 12;12(5):2283–2302. doi: 10.1093/nar/12.5.2283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bendig M. M. Persistence and expression of histone genes injected into Xenopus eggs in early development. Nature. 1981 Jul 2;292(5818):65–67. doi: 10.1038/292065a0. [DOI] [PubMed] [Google Scholar]
  3. Bendig M. M., Williams J. G. Differential expression of the Xenopus laevis tadpole and adult beta-globin genes when injected into fertilized Xenopus laevis eggs. Mol Cell Biol. 1984 Mar;4(3):567–570. doi: 10.1128/mcb.4.3.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bendig M. M., Williams J. G. Replication and expression of Xenopus laevis globin genes injected into fertilized Xenopus eggs. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6197–6201. doi: 10.1073/pnas.80.20.6197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bienz M. Xenopus hsp 70 genes are constitutively expressed in injected oocytes. EMBO J. 1984 Nov;3(11):2477–2483. doi: 10.1002/j.1460-2075.1984.tb02159.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bisbee C. A., Baker M. A., Wilson A. C., Haji-Azimi I., Fischberg M. Albumin phylogeny for clawed frogs (Xenopus). Science. 1977 Feb 25;195(4280):785–787. doi: 10.1126/science.65013. [DOI] [PubMed] [Google Scholar]
  7. Busby S. J., Reeder R. H. Spacer sequences regulate transcription of ribosomal gene plasmids injected into Xenopus embryos. Cell. 1983 Oct;34(3):989–996. doi: 10.1016/0092-8674(83)90556-1. [DOI] [PubMed] [Google Scholar]
  8. Etkin L. D., Balcells S. Transformed Xenopus embryos as a transient expression system to analyze gene expression at the midblastula transition. Dev Biol. 1985 Mar;108(1):173–178. doi: 10.1016/0012-1606(85)90019-3. [DOI] [PubMed] [Google Scholar]
  9. Etkin L. d., Pearman B., Roberts M., Bektesh S. L. Replication, integration and expression of exogenous DNA injected into fertilized eggs of Xenopus laevis. Differentiation. 1984;26(3):194–202. doi: 10.1111/j.1432-0436.1984.tb01395.x. [DOI] [PubMed] [Google Scholar]
  10. Ferguson B., Jones N., Richter J., Rosenberg M. Adenovirus E1a gene product expressed at high levels in Escherichia coli is functional. Science. 1984 Jun 22;224(4655):1343–1346. doi: 10.1126/science.6374895. [DOI] [PubMed] [Google Scholar]
  11. Galli G., Hofstetter H., Stunnenberg H. G., Birnstiel M. L. Biochemical complementation with RNA in the Xenopus oocyte: a small RNA is required for the generation of 3' histone mRNA termini. Cell. 1983 Oct;34(3):823–828. doi: 10.1016/0092-8674(83)90539-1. [DOI] [PubMed] [Google Scholar]
  12. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grichnik J. M., Bergsma D. J., Schwartz R. J. Tissue restricted and stage specific transcription is maintained within 411 nucleotides flanking the 5' end of the chicken alpha-skeletal actin gene. Nucleic Acids Res. 1986 Feb 25;14(4):1683–1701. doi: 10.1093/nar/14.4.1683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gunning P., Mohun T., Ng S. Y., Ponte P., Kedes L. Evolution of the human sarcomeric-actin genes: evidence for units of selection within the 3' untranslated regions of the mRNAs. J Mol Evol. 1984;20(3-4):202–214. doi: 10.1007/BF02104727. [DOI] [PubMed] [Google Scholar]
  15. Gurdon J. B., Fairman S., Mohun T. J., Brennan S. Activation of muscle-specific actin genes in Xenopus development by an induction between animal and vegetal cells of a blastula. Cell. 1985 Jul;41(3):913–922. doi: 10.1016/s0092-8674(85)80072-6. [DOI] [PubMed] [Google Scholar]
  16. Gurdon J. B. Molecular biology in a living cell. Nature. 1974 Apr 26;248(5451):772–776. doi: 10.1038/248772a0. [DOI] [PubMed] [Google Scholar]
  17. Krieg P. A., Melton D. A. Developmental regulation of a gastrula-specific gene injected into fertilized Xenopus eggs. EMBO J. 1985 Dec 16;4(13A):3463–3471. doi: 10.1002/j.1460-2075.1985.tb04105.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mattaj I. W., Lienhard S., Jiricny J., De Robertis E. M. An enhancer-like sequence within the Xenopus U2 gene promoter facilitates the formation of stable transcription complexes. Nature. 1985 Jul 11;316(6024):163–167. doi: 10.1038/316163a0. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Melloul D., Aloni B., Calvo J., Yaffe D., Nudel U. Developmentally regulated expression of chimeric genes containing muscle actin DNA sequences in transfected myogenic cells. EMBO J. 1984 May;3(5):983–990. doi: 10.1002/j.1460-2075.1984.tb01917.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Minty A., Kedes L. Upstream regions of the human cardiac actin gene that modulate its transcription in muscle cells: presence of an evolutionarily conserved repeated motif. Mol Cell Biol. 1986 Jun;6(6):2125–2136. doi: 10.1128/mcb.6.6.2125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mohun T. J., Brennan S., Dathan N., Fairman S., Gurdon J. B. Cell type-specific activation of actin genes in the early amphibian embryo. Nature. 1984 Oct 25;311(5988):716–721. doi: 10.1038/311716a0. [DOI] [PubMed] [Google Scholar]
  24. Newport J., Kirschner M. A major developmental transition in early Xenopus embryos: II. Control of the onset of transcription. Cell. 1982 Oct;30(3):687–696. doi: 10.1016/0092-8674(82)90273-2. [DOI] [PubMed] [Google Scholar]
  25. Nudel U., Greenberg D., Ordahl C. P., Saxel O., Neuman S., Yaffe D. Developmentally regulated expression of a chicken muscle-specific gene in stably transfected rat myogenic cells. Proc Natl Acad Sci U S A. 1985 May;82(10):3106–3109. doi: 10.1073/pnas.82.10.3106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ordahl C. P., Cooper T. A. Strong homology in promoter and 3'-untranslated regions of chick and rat alpha-actin genes. Nature. 1983 May 26;303(5915):348–349. doi: 10.1038/303348a0. [DOI] [PubMed] [Google Scholar]
  27. Rusconi S., Schaffner W. Transformation of frog embryos with a rabbit beta-globin gene. Proc Natl Acad Sci U S A. 1981 Aug;78(8):5051–5055. doi: 10.1073/pnas.78.8.5051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Singh H., Sen R., Baltimore D., Sharp P. A. A nuclear factor that binds to a conserved sequence motif in transcriptional control elements of immunoglobulin genes. Nature. 1986 Jan 9;319(6049):154–158. doi: 10.1038/319154a0. [DOI] [PubMed] [Google Scholar]
  30. Staden R. Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. Nucleic Acids Res. 1982 Aug 11;10(15):4731–4751. doi: 10.1093/nar/10.15.4731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Staden R. Graphic methods to determine the function of nucleic acid sequences. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):521–538. doi: 10.1093/nar/12.1part2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sturgess E. A., Ballantine J. E., Woodland H. R., Mohun P. R., Lane C. D., Dimitriadis G. J. Actin synthesis during the early development of Xenopus laevis. J Embryol Exp Morphol. 1980 Aug;58:303–320. [PubMed] [Google Scholar]
  33. Stutz F., Spohr G. Isolation and characterization of sarcomeric actin genes expressed in Xenopus laevis embryos. J Mol Biol. 1986 Feb 5;187(3):349–361. doi: 10.1016/0022-2836(86)90438-9. [DOI] [PubMed] [Google Scholar]
  34. Zakut R., Shani M., Givol D., Neuman S., Yaffe D., Nudel U. Nucleotide sequence of the rat skeletal muscle actin gene. Nature. 1982 Aug 26;298(5877):857–859. doi: 10.1038/298857a0. [DOI] [PubMed] [Google Scholar]
  35. Zinn K., DiMaio D., Maniatis T. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene. Cell. 1983 Oct;34(3):865–879. doi: 10.1016/0092-8674(83)90544-5. [DOI] [PubMed] [Google Scholar]

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