Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1979 Dec 11;7(7):1749–1763. doi: 10.1093/nar/7.7.1749

A tRNA gene of Xenopus laevis contains at least two sites promoting transcription.

A Kressmann, H Hofstetter, E Di Capua, R Grosschedl, M L Birnstiel
PMCID: PMC342346  PMID: 537910

Abstract

A small cloned DNA segment previously shown to contain all genetic information for the expression of the tRNA1met gene of Xenopus laevis was cleaved into an anterior and posterior portion by Hae III restriction. Both restriction fragments were cloned in pCR1 using EcoRI linkers. Starting from these tDNA subclones, a series of new recombinants were constructed. The transcriptional activity of the cloned DNA's was tested both in an in vitro transcriptional system and by means of the oocyte injection technique. It was shown that both the 5' and 3' ends of the cloned gene unit were essential for transcription. We have developed a model for the functional organization of the tRNA1met gene. We propose that the gene contains a regulatory site situated near the 3' portion of the gene unit. For transcription to occur both in vivo and in vitro a specific initiation site near the 5' end of the gene is required.

Full text

PDF
1749

Images in this article

1755Image
on p.1755
1757Image
on p.1757
1759Image
on p.1759

Selected References

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

  1. Berman M. L., Landy A. Promoter mutations in the transfer RNA gene tyrT of Escherichia coli. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4303–4307. doi: 10.1073/pnas.76.9.4303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birkenmeier E. H., Brown D. D., Jordan E. A nuclear extract of Xenopus laevis oocytes that accurately transcribes 5S RNA genes. Cell. 1978 Nov;15(3):1077–1086. doi: 10.1016/0092-8674(78)90291-x. [DOI] [PubMed] [Google Scholar]
  3. Clarkson S. G., Kurer V., Smith H. O. Sequence organization of a cloned tDNA met fragment from Xenopus laevis. Cell. 1978 Jul;14(3):713–724. doi: 10.1016/0092-8674(78)90253-2. [DOI] [PubMed] [Google Scholar]
  4. Clarkson S. G., Smith H. O., Schaffner W., Gross K. W., Birnstiel M. L. Integration of eukaryotic genes for 5S RNA and histone proteins into a phage lambda receptor. Nucleic Acids Res. 1976 Oct;3(10):2617–2632. doi: 10.1093/nar/3.10.2617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kressmann A., Clarkson S. G., Pirrotta V., Birnstiel M. L. Transcription of cloned tRNA gene fragments and subfragments injected into the oocyte nucleus of Xenopus laevis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1176–1180. doi: 10.1073/pnas.75.3.1176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ng S. Y., Parker C. S., Roeder R. G. Transcription of cloned Xenopus 5S RNA genes by X. laevis RNA polymerase III in reconstituted systems. Proc Natl Acad Sci U S A. 1979 Jan;76(1):136–140. doi: 10.1073/pnas.76.1.136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Schaffner W., Kunz G., Daetwyler H., Telford J., Smith H. O., Birnstiel M. L. Genes and spacers of cloned sea urchin histone DNA analyzed by sequencing. Cell. 1978 Jul;14(3):655–671. doi: 10.1016/0092-8674(78)90249-0. [DOI] [PubMed] [Google Scholar]
  11. Scherer G. E., Walkinshaw M. D., Arnott S. A computer aided oligonucleotide analysis provides a model sequence for RNA polymerase-promoter recognition in E.coli. Nucleic Acids Res. 1978 Oct;5(10):3759–3773. doi: 10.1093/nar/5.10.3759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Telford J. L., Kressmann A., Koski R. A., Grosschedl R., Müller F., Clarkson S. G., Birnstiel M. L. Delimitation of a promoter for RNA polymerase III by means of a functional test. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2590–2594. doi: 10.1073/pnas.76.6.2590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wegnez M., Mazabraud A., Denis H., Pétrissant G., Boisnard M. Biochemical research on oogenesis. Nucleotide sequence of initiator tRNA from oocytes and from somatic cells of Xenopus laevis. Eur J Biochem. 1975 Dec 1;60(1):295–302. doi: 10.1111/j.1432-1033.1975.tb21003.x. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES