The cytoplasm of Xenopus oocytes contains a factor that protects double-stranded RNA from adenosine-to-inosine modification (original) (raw)

Abstract

Here we describe studies of double-stranded RNA (dsRNA) adenosine deaminase in Xenopus laevis, in particular during meiotic maturation, the period during which a stage VI oocyte matures to an egg. We show that dsRNA adenosine deaminase is in the nuclei of stage VI oocytes. Most importantly, we demonstrate that the cytoplasm of stage VI oocytes contains a factor that protects microinjected dsRNA from deamination when dsRNA adenosine deaminase is released from the nucleus during meiotic maturation. Our data suggest that the protection factor is a cytoplasmic dsRNA-binding protein or proteins that bind to dsRNA in a sequence-independent manner to occlude dsRNA from binding to dsRNA adenosine deaminase. The cytoplasmic double-stranded RNA-binding protein(s) does not bind to other nucleic acids and can be titrated at high concentrations of dsRNA. These studies raise the question of whether all dsRNA-binding proteins share endogenous substrates and also suggest potential means of regulating dsRNA adenosine deaminase in vivo.

5425

Images in this article

Selected References

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

  1. Bass B. L., Hurst S. R., Singer J. D. Binding properties of newly identified Xenopus proteins containing dsRNA-binding motifs. Curr Biol. 1994 Apr 1;4(4):301–314. doi: 10.1016/s0960-9822(00)00069-5. [DOI] [PubMed] [Google Scholar]
  2. Bass B. L., Weintraub H. A developmentally regulated activity that unwinds RNA duplexes. Cell. 1987 Feb 27;48(4):607–613. doi: 10.1016/0092-8674(87)90239-x. [DOI] [PubMed] [Google Scholar]
  3. Bass B. L., Weintraub H. An unwinding activity that covalently modifies its double-stranded RNA substrate. Cell. 1988 Dec 23;55(6):1089–1098. doi: 10.1016/0092-8674(88)90253-x. [DOI] [PubMed] [Google Scholar]
  4. Chang H. W., Watson J. C., Jacobs B. L. The E3L gene of vaccinia virus encodes an inhibitor of the interferon-induced, double-stranded RNA-dependent protein kinase. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4825–4829. doi: 10.1073/pnas.89.11.4825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feldherr C. M., Richmond P. A. Manual enucleation of Xenopus oocytes. Methods Cell Biol. 1978;17:75–79. doi: 10.1016/s0091-679x(08)61135-8. [DOI] [PubMed] [Google Scholar]
  6. Fu L. N., Ye R. Q., Browder L. W., Johnston R. N. Translational potentiation of messenger RNA with secondary structure in Xenopus. Science. 1991 Feb 15;251(4995):807–810. doi: 10.1126/science.1990443. [DOI] [PubMed] [Google Scholar]
  7. Gatignol A., Buckler C., Jeang K. T. Relatedness of an RNA-binding motif in human immunodeficiency virus type 1 TAR RNA-binding protein TRBP to human P1/dsI kinase and Drosophila staufen. Mol Cell Biol. 1993 Apr;13(4):2193–2202. doi: 10.1128/mcb.13.4.2193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Harland R., Weintraub H. Translation of mRNA injected into Xenopus oocytes is specifically inhibited by antisense RNA. J Cell Biol. 1985 Sep;101(3):1094–1099. doi: 10.1083/jcb.101.3.1094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Higuchi M., Single F. N., Köhler M., Sommer B., Sprengel R., Seeburg P. H. RNA editing of AMPA receptor subunit GluR-B: a base-paired intron-exon structure determines position and efficiency. Cell. 1993 Dec 31;75(7):1361–1370. doi: 10.1016/0092-8674(93)90622-w. [DOI] [PubMed] [Google Scholar]
  10. Hough R. F., Bass B. L. Purification of the Xenopus laevis double-stranded RNA adenosine deaminase. J Biol Chem. 1994 Apr 1;269(13):9933–9939. [PubMed] [Google Scholar]
  11. Imani F., Jacobs B. L. Inhibitory activity for the interferon-induced protein kinase is associated with the reovirus serotype 1 sigma 3 protein. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7887–7891. doi: 10.1073/pnas.85.21.7887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kim U., Nishikura K. Double-stranded RNA adenosine deaminase as a potential mammalian RNA editing factor. Semin Cell Biol. 1993 Aug;4(4):285–293. doi: 10.1006/scel.1993.1034. [DOI] [PubMed] [Google Scholar]
  13. Kimelman D., Kirschner M. W. An antisense mRNA directs the covalent modification of the transcript encoding fibroblast growth factor in Xenopus oocytes. Cell. 1989 Nov 17;59(4):687–696. doi: 10.1016/0092-8674(89)90015-9. [DOI] [PubMed] [Google Scholar]
  14. Lloyd R. M., Shatkin A. J. Translational stimulation by reovirus polypeptide sigma 3: substitution for VAI RNA and inhibition of phosphorylation of the alpha subunit of eukaryotic initiation factor 2. J Virol. 1992 Dec;66(12):6878–6884. doi: 10.1128/jvi.66.12.6878-6884.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Milligan J. F., Groebe D. R., Witherell G. W., Uhlenbeck O. C. Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates. Nucleic Acids Res. 1987 Nov 11;15(21):8783–8798. doi: 10.1093/nar/15.21.8783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nishikura K., Yoo C., Kim U., Murray J. M., Estes P. A., Cash F. E., Liebhaber S. A. Substrate specificity of the dsRNA unwinding/modifying activity. EMBO J. 1991 Nov;10(11):3523–3532. doi: 10.1002/j.1460-2075.1991.tb04916.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Polson A. G., Crain P. F., Pomerantz S. C., McCloskey J. A., Bass B. L. The mechanism of adenosine to inosine conversion by the double-stranded RNA unwinding/modifying activity: a high-performance liquid chromatography-mass spectrometry analysis. Biochemistry. 1991 Dec 10;30(49):11507–11514. doi: 10.1021/bi00113a004. [DOI] [PubMed] [Google Scholar]
  18. Rebagliati M. R., Melton D. A. Antisense RNA injections in fertilized frog eggs reveal an RNA duplex unwinding activity. Cell. 1987 Feb 27;48(4):599–605. doi: 10.1016/0092-8674(87)90238-8. [DOI] [PubMed] [Google Scholar]
  19. Samuel C. E. Antiviral actions of interferon. Interferon-regulated cellular proteins and their surprisingly selective antiviral activities. Virology. 1991 Jul;183(1):1–11. doi: 10.1016/0042-6822(91)90112-o. [DOI] [PubMed] [Google Scholar]
  20. Sharmeen L., Bass B., Sonenberg N., Weintraub H., Groudine M. Tat-dependent adenosine-to-inosine modification of wild-type transactivation response RNA. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8096–8100. doi: 10.1073/pnas.88.18.8096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Skeiky Y. A., Iatrou K. Developmental regulation of covalent modification of double-stranded RNA during silkmoth oogenesis. J Mol Biol. 1991 Apr 5;218(3):517–527. doi: 10.1016/0022-2836(91)90698-6. [DOI] [PubMed] [Google Scholar]
  22. Smith L. D. The induction of oocyte maturation: transmembrane signaling events and regulation of the cell cycle. Development. 1989 Dec;107(4):685–699. doi: 10.1242/dev.107.4.685. [DOI] [PubMed] [Google Scholar]
  23. St Johnston D., Brown N. H., Gall J. G., Jantsch M. A conserved double-stranded RNA-binding domain. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10979–10983. doi: 10.1073/pnas.89.22.10979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wagner R. W., Nishikura K. Cell cycle expression of RNA duplex unwindase activity in mammalian cells. Mol Cell Biol. 1988 Feb;8(2):770–777. doi: 10.1128/mcb.8.2.770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wagner R. W., Yoo C., Wrabetz L., Kamholz J., Buchhalter J., Hassan N. F., Khalili K., Kim S. U., Perussia B., McMorris F. A. Double-stranded RNA unwinding and modifying activity is detected ubiquitously in primary tissues and cell lines. Mol Cell Biol. 1990 Oct;10(10):5586–5590. doi: 10.1128/mcb.10.10.5586. [DOI] [PMC free article] [PubMed] [Google Scholar]