Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1997 May;17(5):2413–2424. doi: 10.1128/mcb.17.5.2413

Editing of glutamate receptor B subunit ion channel RNAs by four alternatively spliced DRADA2 double-stranded RNA adenosine deaminases.

F Lai 1, C X Chen 1, K C Carter 1, K Nishikura 1
PMCID: PMC232090  PMID: 9111310

Abstract

Double-stranded (ds) RNA-specific adenosine deaminase converts adenosine residues into inosines in dsRNA and edits transcripts of certain cellular and viral genes such as glutamate receptor (GluR) subunits and hepatitis delta antigen. The first member of this type of deaminase, DRADA1, has been recently cloned based on the amino acid sequence information derived from biochemically purified proteins. Our search for DRADA1-like genes through expressed sequence tag databases led to the cloning of the second member of this class of enzyme, DRADA2, which has a high degree of sequence homology to DRADA1 yet exhibits a distinctive RNA editing site selectivity. There are four differentially spliced isoforms of human DRADA2. These different isoforms of recombinant DRADA2 proteins, including one which is a human homolog of the recently reported rat RED1, were analyzed in vitro for their GluR B subunit (GluR-B) RNA editing site selectivity. As originally reported for rat RED1, the DRADA2a and -2b isoforms edit GluR-B RNA efficiently at the so-called Q/R site, whereas DRADA1 barely edits this site. In contrast, the R/G site of GluR-B RNA was edited efficiently by the DRADA2a and -2b isoforms as well as DRADA1. Isoforms DRADA2c and -2d, which have a distinctive truncated shorter C-terminal structure, displayed weak adenosine-to-inosine conversion activity but no editing activity tested at three known sites of GluR-B RNA. The possible role of these DRADA2c and -2d isoforms in the regulatory mechanism of RNA editing is discussed.

Full Text

The Full Text of this article is available as a PDF (758.4 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bass B. L. RNA editing. An I for editing. Curr Biol. 1995 Jun 1;5(6):598–600. doi: 10.1016/s0960-9822(95)00119-9. [DOI] [PubMed] [Google Scholar]
  3. Benne R. RNA editing. The long and the short of it. Nature. 1996 Apr 4;380(6573):391–392. doi: 10.1038/380391a0. [DOI] [PubMed] [Google Scholar]
  4. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bernard A., Khrestchatisky M. Assessing the extent of RNA editing in the TMII regions of GluR5 and GluR6 kainate receptors during rat brain development. J Neurochem. 1994 May;62(5):2057–2060. doi: 10.1046/j.1471-4159.1994.62052057.x. [DOI] [PubMed] [Google Scholar]
  6. Burnashev N., Monyer H., Seeburg P. H., Sakmann B. Divalent ion permeability of AMPA receptor channels is dominated by the edited form of a single subunit. Neuron. 1992 Jan;8(1):189–198. doi: 10.1016/0896-6273(92)90120-3. [DOI] [PubMed] [Google Scholar]
  7. Caras I. W., Davitz M. A., Rhee L., Weddell G., Martin D. W., Jr, Nussenzweig V. Cloning of decay-accelerating factor suggests novel use of splicing to generate two proteins. Nature. 1987 Feb 5;325(6104):545–549. doi: 10.1038/325545a0. [DOI] [PubMed] [Google Scholar]
  8. Cheng J. F., Boyartchuk V., Zhu Y. Isolation and mapping of human chromosome 21 cDNA: progress in constructing a chromosome 21 expression map. Genomics. 1994 Sep 1;23(1):75–84. doi: 10.1006/geno.1994.1461. [DOI] [PubMed] [Google Scholar]
  9. Dabiri G. A., Lai F., Drakas R. A., Nishikura K. Editing of the GLuR-B ion channel RNA in vitro by recombinant double-stranded RNA adenosine deaminase. EMBO J. 1996 Jan 2;15(1):34–45. [PMC free article] [PubMed] [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Egebjerg J., Kukekov V., Heinemann S. F. Intron sequence directs RNA editing of the glutamate receptor subunit GluR2 coding sequence. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10270–10274. doi: 10.1073/pnas.91.22.10270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Görlich D., Mattaj I. W. Nucleocytoplasmic transport. Science. 1996 Mar 15;271(5255):1513–1518. doi: 10.1126/science.271.5255.1513. [DOI] [PubMed] [Google Scholar]
  13. Herb A., Higuchi M., Sprengel R., Seeburg P. H. Q/R site editing in kainate receptor GluR5 and GluR6 pre-mRNAs requires distant intronic sequences. Proc Natl Acad Sci U S A. 1996 Mar 5;93(5):1875–1880. doi: 10.1073/pnas.93.5.1875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Kao P. N., Chen L., Brock G., Ng J., Kenny J., Smith A. J., Corthésy B. Cloning and expression of cyclosporin A- and FK506-sensitive nuclear factor of activated T-cells: NF45 and NF90. J Biol Chem. 1994 Aug 12;269(32):20691–20699. [PubMed] [Google Scholar]
  16. Kim U., Garner T. L., Sanford T., Speicher D., Murray J. M., Nishikura K. Purification and characterization of double-stranded RNA adenosine deaminase from bovine nuclear extracts. J Biol Chem. 1994 May 6;269(18):13480–13489. [PubMed] [Google Scholar]
  17. Kim U., Wang Y., Sanford T., Zeng Y., Nishikura K. Molecular cloning of cDNA for double-stranded RNA adenosine deaminase, a candidate enzyme for nuclear RNA editing. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11457–11461. doi: 10.1073/pnas.91.24.11457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Korenberg J. R., Chen X. N., Schipper R., Sun Z., Gonsky R., Gerwehr S., Carpenter N., Daumer C., Dignan P., Disteche C. Down syndrome phenotypes: the consequences of chromosomal imbalance. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4997–5001. doi: 10.1073/pnas.91.11.4997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Köhler M., Burnashev N., Sakmann B., Seeburg P. H. Determinants of Ca2+ permeability in both TM1 and TM2 of high affinity kainate receptor channels: diversity by RNA editing. Neuron. 1993 Mar;10(3):491–500. doi: 10.1016/0896-6273(93)90336-p. [DOI] [PubMed] [Google Scholar]
  21. Köhler M., Kornau H. C., Seeburg P. H. The organization of the gene for the functionally dominant alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor subunit GluR-B. J Biol Chem. 1994 Jul 1;269(26):17367–17370. [PubMed] [Google Scholar]
  22. Lai F., Drakas R., Nishikura K. Mutagenic analysis of double-stranded RNA adenosine deaminase, a candidate enzyme for RNA editing of glutamate-gated ion channel transcripts. J Biol Chem. 1995 Jul 21;270(29):17098–17105. doi: 10.1074/jbc.270.29.17098. [DOI] [PubMed] [Google Scholar]
  23. Lomeli H., Mosbacher J., Melcher T., Höger T., Geiger J. R., Kuner T., Monyer H., Higuchi M., Bach A., Seeburg P. H. Control of kinetic properties of AMPA receptor channels by nuclear RNA editing. Science. 1994 Dec 9;266(5191):1709–1713. doi: 10.1126/science.7992055. [DOI] [PubMed] [Google Scholar]
  24. Maas S., Melcher T., Herb A., Seeburg P. H., Keller W., Krause S., Higuchi M., O'Connell M. A. Structural requirements for RNA editing in glutamate receptor pre-mRNAs by recombinant double-stranded RNA adenosine deaminase. J Biol Chem. 1996 May 24;271(21):12221–12226. doi: 10.1074/jbc.271.21.12221. [DOI] [PubMed] [Google Scholar]
  25. Makałowski W., Mitchell G. A., Labuda D. Alu sequences in the coding regions of mRNA: a source of protein variability. Trends Genet. 1994 Jun;10(6):188–193. doi: 10.1016/0168-9525(94)90254-2. [DOI] [PubMed] [Google Scholar]
  26. Melcher T., Maas S., Herb A., Sprengel R., Seeburg P. H., Higuchi M. A mammalian RNA editing enzyme. Nature. 1996 Feb 1;379(6564):460–464. doi: 10.1038/379460a0. [DOI] [PubMed] [Google Scholar]
  27. Nutt S. L., Kamboj R. K. Differential RNA editing efficiency of AMPA receptor subunit GluR-2 in human brain. Neuroreport. 1994 Aug 15;5(13):1679–1683. doi: 10.1097/00001756-199408150-00034. [DOI] [PubMed] [Google Scholar]
  28. O'Connell M. A., Krause S., Higuchi M., Hsuan J. J., Totty N. F., Jenny A., Keller W. Cloning of cDNAs encoding mammalian double-stranded RNA-specific adenosine deaminase. Mol Cell Biol. 1995 Mar;15(3):1389–1397. doi: 10.1128/mcb.15.3.1389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Paschen W., Djuricic B. Regional differences in the extent of RNA editing of the glutamate receptor subunits GluR2 and GluR6 in rat brain. J Neurosci Methods. 1995 Jan;56(1):21–29. doi: 10.1016/0165-0270(94)00085-u. [DOI] [PubMed] [Google Scholar]
  30. Patterson J. B., Samuel C. E. Expression and regulation by interferon of a double-stranded-RNA-specific adenosine deaminase from human cells: evidence for two forms of the deaminase. Mol Cell Biol. 1995 Oct;15(10):5376–5388. doi: 10.1128/mcb.15.10.5376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  32. Puchalski R. B., Louis J. C., Brose N., Traynelis S. F., Egebjerg J., Kukekov V., Wenthold R. J., Rogers S. W., Lin F., Moran T. Selective RNA editing and subunit assembly of native glutamate receptors. Neuron. 1994 Jul;13(1):131–147. doi: 10.1016/0896-6273(94)90464-2. [DOI] [PubMed] [Google Scholar]
  33. Ruano D., Lambolez B., Rossier J., Paternain A. V., Lerma J. Kainate receptor subunits expressed in single cultured hippocampal neurons: molecular and functional variants by RNA editing. Neuron. 1995 May;14(5):1009–1017. doi: 10.1016/0896-6273(95)90339-9. [DOI] [PubMed] [Google Scholar]
  34. Rueter S. M., Burns C. M., Coode S. A., Mookherjee P., Emeson R. B. Glutamate receptor RNA editing in vitro by enzymatic conversion of adenosine to inosine. Science. 1995 Mar 10;267(5203):1491–1494. doi: 10.1126/science.7878468. [DOI] [PubMed] [Google Scholar]
  35. Schumacher J. M., Lee K., Edelhoff S., Braun R. E. Spnr, a murine RNA-binding protein that is localized to cytoplasmic microtubules. J Cell Biol. 1995 May;129(4):1023–1032. doi: 10.1083/jcb.129.4.1023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Scott J. A place in the world for RNA editing. Cell. 1995 Jun 16;81(6):833–836. doi: 10.1016/0092-8674(95)90002-0. [DOI] [PubMed] [Google Scholar]
  37. Seeburg P. H. The role of RNA editing in controlling glutamate receptor channel properties. J Neurochem. 1996 Jan;66(1):1–5. doi: 10.1046/j.1471-4159.1996.66010001.x. [DOI] [PubMed] [Google Scholar]
  38. Sommer B., Köhler M., Sprengel R., Seeburg P. H. RNA editing in brain controls a determinant of ion flow in glutamate-gated channels. Cell. 1991 Oct 4;67(1):11–19. doi: 10.1016/0092-8674(91)90568-j. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Verdoorn T. A., Burnashev N., Monyer H., Seeburg P. H., Sakmann B. Structural determinants of ion flow through recombinant glutamate receptor channels. Science. 1991 Jun 21;252(5013):1715–1718. doi: 10.1126/science.1710829. [DOI] [PubMed] [Google Scholar]
  41. Wang Y., Zeng Y., Murray J. M., Nishikura K. Genomic organization and chromosomal location of the human dsRNA adenosine deaminase gene: the enzyme for glutamate-activated ion channel RNA editing. J Mol Biol. 1995 Nov 24;254(2):184–195. doi: 10.1006/jmbi.1995.0610. [DOI] [PubMed] [Google Scholar]
  42. Weier H. U., George C. X., Greulich K. M., Samuel C. E. The interferon-inducible, double-stranded RNA-specific adenosine deaminase gene (DSRAD) maps to human chromosome 1q21.1-21.2. Genomics. 1995 Nov 20;30(2):372–375. doi: 10.1006/geno.1995.0034. [DOI] [PubMed] [Google Scholar]
  43. Yang J. H., Sklar P., Axel R., Maniatis T. Editing of glutamate receptor subunit B pre-mRNA in vitro by site-specific deamination of adenosine. Nature. 1995 Mar 2;374(6517):77–81. doi: 10.1038/374077a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES