Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1982 Mar;2(3):241–249. doi: 10.1128/mcb.2.3.241

Nuclear ligation of RNA 5'-OH kinase products in tRNA.

I Winicov, J D Button
PMCID: PMC369782  PMID: 7110132

Abstract

Mouse L-cell nuclei incorporate gamma-32P from ATP in vitro predominantly in 5'-monophosphoryl termini and internal phosphodiester bonds with a nonrandom nearest-neighbor distribution. In the presence of 1 microgram of alpha-amanitin per ml the gamma-32P showed a time-dependent appearance in RNA bands which migrated with mature tRNA species but not with pre-tRNA and 5S RNA. The gamma-32P was found in internal phosphodiester bonds as shown by alkaline phosphatase resistance and was identified in 3'-monophosphates after RNase T2, T1, and A digestion. The specificity of this incorporation was indicated by a limited number of labeled oligonucleotides from a T1 digest and identification of 70 to 80% of the 32P label as Cp on complete digestion of the eluted tRNA band. We also observed transiently [gamma-32P]ATP-labeled RNA bands (in 5'-monophosphate positions) that were 32 to 45 nucleotides long. The results presented suggest splicing of several mouse L-cell tRNA species in isolated nuclei which involve the RNA 5'-OH kinase products as intermediates.

Full text

PDF
241

Images in this article

Selected References

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

  1. Abelson J. RNA processing and the intervening sequence problem. Annu Rev Biochem. 1979;48:1035–1069. doi: 10.1146/annurev.bi.48.070179.005131. [DOI] [PubMed] [Google Scholar]
  2. Altman S. Transfer RNA processing enzymes. Cell. 1981 Jan;23(1):3–4. doi: 10.1016/0092-8674(81)90262-2. [DOI] [PubMed] [Google Scholar]
  3. Carin M., Jensen B. F., Jentsch K. D., Leer J. C., Nielsen O. F., Westergaard O. In vitro splicing of the ribosomal RNA precursor in isolated nucleoli from Tetrahymena. Nucleic Acids Res. 1980 Dec 11;8(23):5551–5566. doi: 10.1093/nar/8.23.5551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Contreras R., Fiers W. Initiation of transcription by RNA polymerase II in permeable, SV40-infected or noninfected, CVI cells; evidence for multiple promoters of SV40 late transcription. Nucleic Acids Res. 1981 Jan 24;9(2):215–236. doi: 10.1093/nar/9.2.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. De Robertis E. M., Black P., Nishikura K. Intranuclear location of the tRNA splicing enzymes. Cell. 1981 Jan;23(1):89–93. doi: 10.1016/0092-8674(81)90273-7. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Deutscher M. P. Synthesis and functions of the -C-C-A terminus of transfer RNA. Prog Nucleic Acid Res Mol Biol. 1973;13:51–92. doi: 10.1016/s0079-6603(08)60100-2. [DOI] [PubMed] [Google Scholar]
  8. Etcheverry T., Colby D., Guthrie C. A precursor to a minor species of yeast tRNASer contains an intervening sequence. Cell. 1979 Sep;18(1):11–26. doi: 10.1016/0092-8674(79)90349-0. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Gorecki M., Wilchek M., Patchornik A. The conversion of 3-monoazotyrosine to 3-aminotyrosine in peptides and proteins. Biochim Biophys Acta. 1971 Mar 23;229(3):590–595. doi: 10.1016/0005-2795(71)90274-1. [DOI] [PubMed] [Google Scholar]
  11. Knapp G., Beckmann J. S., Johnson P. F., Fuhrman S. A., Abelson J. Transcription and processing of intervening sequences in yeast tRNA genes. Cell. 1978 Jun;14(2):221–236. doi: 10.1016/0092-8674(78)90109-5. [DOI] [PubMed] [Google Scholar]
  12. Knapp G., Ogden R. C., Peebles C. L., Abelson J. Splicing of yeast tRNA precursors: structure of the reaction intermediates. Cell. 1979 Sep;18(1):37–45. doi: 10.1016/0092-8674(79)90351-9. [DOI] [PubMed] [Google Scholar]
  13. Konarska M., Filipowicz W., Domdey H., Gross H. J. Formation of a 2'-phosphomonoester, 3',5'-phosphodiester linkage by a novel RNA ligase in wheat germ. Nature. 1981 Sep 10;293(5828):112–116. doi: 10.1038/293112a0. [DOI] [PubMed] [Google Scholar]
  14. Lönn U. Exclusive nuclear location of precursor 4 S RNA in vivo. J Mol Biol. 1977 Jun 5;112(4):661–666. doi: 10.1016/s0022-2836(77)80172-1. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. O'Farrell P. Z., Cordell B., Valenzuela P., Rutter W. J., Goodman H. M. Structure and processing of yeast precursor tRNAs containing intervening sequences. Nature. 1978 Aug 3;274(5670):438–445. doi: 10.1038/274438a0. [DOI] [PubMed] [Google Scholar]
  17. Ogden R. C., Beckman J. S., Abelson J., Kang H. S., Söll D., Schmidt O. In vitro transcription and processing of a yeast tRNA gene containing an intervening sequence. Cell. 1979 Jun;17(2):399–406. doi: 10.1016/0092-8674(79)90166-1. [DOI] [PubMed] [Google Scholar]
  18. Otsuka A., de Paolis A., Tocchini-Valentini G. P. Ribonuclease "XlaI," an activity from Xenopus laevis oocytes that excises intervening sequences from yeast transfer ribonucleic acid precursors. Mol Cell Biol. 1981 Mar;1(3):269–280. doi: 10.1128/mcb.1.3.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Peebles C. L., Ogden R. C., Knapp G., Abelson J. Splicing of yeast tRNA precursors: a two-stage reaction. Cell. 1979 Sep;18(1):27–35. doi: 10.1016/0092-8674(79)90350-7. [DOI] [PubMed] [Google Scholar]
  20. Robinson R. R., Davidson N. Analysis of a drosophila tRNA gene cluster: two tRNALeu genes contain intervening sequences. Cell. 1981 Jan;23(1):251–259. doi: 10.1016/0092-8674(81)90289-0. [DOI] [PubMed] [Google Scholar]
  21. Valenzuela P., Venegas A., Weinberg F., Bishop R., Rutter W. J. Structure of yeast phenylalanine-tRNA genes: an intervening DNA segment within the region coding for the tRNA. Proc Natl Acad Sci U S A. 1978 Jan;75(1):190–194. doi: 10.1073/pnas.75.1.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Winicov I. Nuclear transcription in vitro. Sensitivity to inhibition by ribosyldichlorobenzimidazole and rifamycin AF/013. Biochemistry. 1979 Apr 17;18(8):1575–1582. doi: 10.1021/bi00575a030. [DOI] [PubMed] [Google Scholar]
  23. Winicov I., Perry R. P. Synthesis methylation, and capping of nuclear RNA by a subcellular system. Biochemistry. 1976 Nov 16;15(23):5039–5046. doi: 10.1021/bi00668a014. [DOI] [PubMed] [Google Scholar]
  24. Winicov I. RNA phosphorylation: a polynucleotide kinase function in mouse L cell nuclei. Biochemistry. 1977 Sep 20;16(19):4233–4237. doi: 10.1021/bi00638a016. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES