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
. 1980 Jul;77(7):4147–4151. doi: 10.1073/pnas.77.7.4147

Transcription of tRNA genes in vivo: single-stranded compared to double-stranded templates.

R Cortese, R Harland, D Melton
PMCID: PMC349787  PMID: 7001455

Abstract

The expression of cloned tRNA genes has been studied by injecting single-stranded and double-stranded DNA templates into Xenopus oocyte nuclei. In both forms the genes are faithfully transcribed after injection. Some single-stranded DNA is converted into double-stranded DNA in the oocyte nucleus. This conversion is necessary for the expression of the injected tRNA gene: no tRNA transcription is observed when DNA synthesis is inhibited. We conclude that single-stranded DNA does not serve as a template for faithful transcription of this gene in injected oocytes.

Full text

PDF
4147

Images in this article

Selected References

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

  1. Brown D. D., Gurdon J. B. High-fidelity transcription of 5S DNA injected into Xenopus oocytes. Proc Natl Acad Sci U S A. 1977 May;74(5):2064–2068. doi: 10.1073/pnas.74.5.2064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cortese R., Melton D., Tranquilla T., Smith J. D. Cloning of nematode tRNA genes and their expression in the frog oocyte. Nucleic Acids Res. 1978 Dec;5(12):4593–4611. [PMC free article] [PubMed] [Google Scholar]
  4. De Robertis E. M., Mertz J. E. Coupled transcription-translation of DNA injected into Xenopus oocytes. Cell. 1977 Sep;12(1):175–182. doi: 10.1016/0092-8674(77)90195-7. [DOI] [PubMed] [Google Scholar]
  5. Gurdon J. B. Injected nuclei in frog oocytes: fate, enlargement, and chromatin dispersal. J Embryol Exp Morphol. 1976 Dec;36(3):523–540. [PubMed] [Google Scholar]
  6. Gurdon J. B., Lane C. D., Woodland H. R., Marbaix G. Use of frog eggs and oocytes for the study of messenger RNA and its translation in living cells. Nature. 1971 Sep 17;233(5316):177–182. doi: 10.1038/233177a0. [DOI] [PubMed] [Google Scholar]
  7. Hunter T., Francke B. In vitro polyoma DNA synthesis: inhibition by 1-beta-d-arabinofuranosyl CTP. J Virol. 1975 Apr;15(4):759–775. doi: 10.1128/jvi.15.4.759-775.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Lilley D. M., Houghton M. The interaction of RNA polymerase II from wheat with supercoiled and linear plasmid templates. Nucleic Acids Res. 1979 Feb;6(2):507–523. doi: 10.1093/nar/6.2.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Melton D. A., Cortese R. Transcription of cloned tRNA genes and the nuclear partitioning of a tRNA precursor. Cell. 1979 Dec;18(4):1165–1172. doi: 10.1016/0092-8674(79)90229-0. [DOI] [PubMed] [Google Scholar]
  11. Melton D. A., De Robertis E. M., Cortese R. Order and intracellular location of the events involved in the maturation of a spliced tRNA. Nature. 1980 Mar 13;284(5752):143–148. doi: 10.1038/284143a0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  14. Schreier P. H., Cortese R. A fast and simple method for sequencing DNA cloned in the single-stranded bacteriophage M13. J Mol Biol. 1979 Mar 25;129(1):169–172. doi: 10.1016/0022-2836(79)90068-8. [DOI] [PubMed] [Google Scholar]
  15. 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]

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