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. 1998 Feb;4(2):195–204.

The U14 snoRNA is required for 2'-O-methylation of the pre-18S rRNA in Xenopus oocytes.

D A Dunbar 1, S J Baserga 1
PMCID: PMC1369608  PMID: 9570319

Abstract

We have studied the role of the U14 small nucleolar RNA (snoRNA) in pre-rRNA methylation and processing in Xenopus oocytes. Depletion of U14 in Xenopus oocytes was achieved by co-injecting two nonoverlapping antisense oligonucleotides. Focusing on the earliest precursor, depletion experiments revealed that the U14 snoRNA is essential for 2'-O-ribose methylation at nt 427 of the 18S rRNA. Injection of U14-depleted oocytes with specific U14 mutant snoRNAs indicated that conserved domain B, but not domain A, of U14 is required for the methylation reaction. When the effect of U14 on pre-rRNA processing is assayed, we find only modest effects on 18S rRNA levels, and no effect on the type or accumulation of 18S precursors, suggesting a role for U14 in a step in ribosome biogenesis other than cleavage of the pre-rRNA. Xenopus U14 is, therefore, a Box C/D fibrillarin-associated snoRNA that is required for site-specific 2'-O-ribose methylation of pre-rRNA.

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Selected References

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  1. Anderson D. M., Smith L. D. Synthesis of heterogeneous nuclear RNA in full-grown oocytes of Xenopus laevis (Daudin). Cell. 1977 Jul;11(3):663–671. doi: 10.1016/0092-8674(77)90083-6. [DOI] [PubMed] [Google Scholar]
  2. Balakin A. G., Smith L., Fournier M. J. The RNA world of the nucleolus: two major families of small RNAs defined by different box elements with related functions. Cell. 1996 Sep 6;86(5):823–834. doi: 10.1016/s0092-8674(00)80156-7. [DOI] [PubMed] [Google Scholar]
  3. Baserga S. J., Yang X. D., Steitz J. A. An intact Box C sequence in the U3 snRNA is required for binding of fibrillarin, the protein common to the major family of nucleolar snRNPs. EMBO J. 1991 Sep;10(9):2645–2651. doi: 10.1002/j.1460-2075.1991.tb07807.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cavaillé J., Nicoloso M., Bachellerie J. P. Targeted ribose methylation of RNA in vivo directed by tailored antisense RNA guides. Nature. 1996 Oct 24;383(6602):732–735. doi: 10.1038/383732a0. [DOI] [PubMed] [Google Scholar]
  5. Dunbar D. A., Ware V. C., Baserga S. J. The U18 snRNA is not essential for pre-rRNA processing in Xenopus laevis. RNA. 1996 Apr;2(4):324–333. [PMC free article] [PubMed] [Google Scholar]
  6. Enright C. A., Maxwell E. S., Eliceiri G. L., Sollner-Webb B. 5'ETS rRNA processing facilitated by four small RNAs: U14, E3, U17, and U3. RNA. 1996 Nov;2(11):1094–1099. [PMC free article] [PubMed] [Google Scholar]
  7. Ganot P., Bortolin M. L., Kiss T. Site-specific pseudouridine formation in preribosomal RNA is guided by small nucleolar RNAs. Cell. 1997 May 30;89(5):799–809. doi: 10.1016/s0092-8674(00)80263-9. [DOI] [PubMed] [Google Scholar]
  8. Hughes J. M., Ares M., Jr Depletion of U3 small nucleolar RNA inhibits cleavage in the 5' external transcribed spacer of yeast pre-ribosomal RNA and impairs formation of 18S ribosomal RNA. EMBO J. 1991 Dec;10(13):4231–4239. doi: 10.1002/j.1460-2075.1991.tb05001.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Inoue H., Hayase Y., Iwai S., Ohtsuka E. Sequence-dependent hydrolysis of RNA using modified oligonucleotide splints and RNase H. FEBS Lett. 1987 May 11;215(2):327–330. doi: 10.1016/0014-5793(87)80171-0. [DOI] [PubMed] [Google Scholar]
  10. Kass S., Tyc K., Steitz J. A., Sollner-Webb B. The U3 small nucleolar ribonucleoprotein functions in the first step of preribosomal RNA processing. Cell. 1990 Mar 23;60(6):897–908. doi: 10.1016/0092-8674(90)90338-f. [DOI] [PubMed] [Google Scholar]
  11. Kiss-László Z., Henry Y., Bachellerie J. P., Caizergues-Ferrer M., Kiss T. Site-specific ribose methylation of preribosomal RNA: a novel function for small nucleolar RNAs. Cell. 1996 Jun 28;85(7):1077–1088. doi: 10.1016/s0092-8674(00)81308-2. [DOI] [PubMed] [Google Scholar]
  12. Kumar A. Ribosome synthesis in Tetrahymena pyriformis. J Cell Biol. 1970 Jun;45(3):623–634. doi: 10.1083/jcb.45.3.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lapham J., Crothers D. M. RNase H cleavage for processing of in vitro transcribed RNA for NMR studies and RNA ligation. RNA. 1996 Mar;2(3):289–296. [PMC free article] [PubMed] [Google Scholar]
  14. Lapham J., Yu Y. T., Shu M. D., Steitz J. A., Crothers D. M. The position of site-directed cleavage of RNA using RNase H and 2'-O-methyl oligonucleotides is dependent on the enzyme source. RNA. 1997 Sep;3(9):950–951. [PMC free article] [PubMed] [Google Scholar]
  15. Leader D. J., Sanders J. F., Waugh R., Shaw P., Brown J. W. Molecular characterisation of plant U14 small nucleolar RNA genes: closely linked genes are transcribed as polycistronic U14 transcripts. Nucleic Acids Res. 1994 Dec 11;22(24):5196–5203. doi: 10.1093/nar/22.24.5196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Li D., Fournier M. J. U14 function in Saccharomyces cerevisiae can be provided by large deletion variants of yeast U14 and hybrid mouse-yeast U14 RNAs. EMBO J. 1992 Feb;11(2):683–689. doi: 10.1002/j.1460-2075.1992.tb05100.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Li H. D., Zagorski J., Fournier M. J. Depletion of U14 small nuclear RNA (snR128) disrupts production of 18S rRNA in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Mar;10(3):1145–1152. doi: 10.1128/mcb.10.3.1145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liang W. Q., Clark J. A., Fournier M. J. The rRNA-processing function of the yeast U14 small nucleolar RNA can be rescued by a conserved RNA helicase-like protein. Mol Cell Biol. 1997 Jul;17(7):4124–4132. doi: 10.1128/mcb.17.7.4124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Liang W. Q., Fournier M. J. U14 base-pairs with 18S rRNA: a novel snoRNA interaction required for rRNA processing. Genes Dev. 1995 Oct 1;9(19):2433–2443. doi: 10.1101/gad.9.19.2433. [DOI] [PubMed] [Google Scholar]
  20. Lindahl L., Archer R. H., Zengel J. M. A new rRNA processing mutant of Saccharomyces cerevisiae. Nucleic Acids Res. 1992 Jan 25;20(2):295–301. doi: 10.1093/nar/20.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Liu J., Maxwell E. S. Mouse U14 snRNA is encoded in an intron of the mouse cognate hsc70 heat shock gene. Nucleic Acids Res. 1990 Nov 25;18(22):6565–6571. doi: 10.1093/nar/18.22.6565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maden B. E., Hughes J. M. Eukaryotic ribosomal RNA: the recent excitement in the nucleotide modification problem. Chromosoma. 1997 Jun;105(7-8):391–400. doi: 10.1007/BF02510475. [DOI] [PubMed] [Google Scholar]
  23. Maden B. E. The numerous modified nucleotides in eukaryotic ribosomal RNA. Prog Nucleic Acid Res Mol Biol. 1990;39:241–303. doi: 10.1016/s0079-6603(08)60629-7. [DOI] [PubMed] [Google Scholar]
  24. Maxwell E. S., Fournier M. J. The small nucleolar RNAs. Annu Rev Biochem. 1995;64:897–934. doi: 10.1146/annurev.bi.64.070195.004341. [DOI] [PubMed] [Google Scholar]
  25. Mishra R. K., Eliceiri G. L. Three small nucleolar RNAs that are involved in ribosomal RNA precursor processing. Proc Natl Acad Sci U S A. 1997 May 13;94(10):4972–4977. doi: 10.1073/pnas.94.10.4972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morrissey J. P., Tollervey D. U14 small nucleolar RNA makes multiple contacts with the pre-ribosomal RNA. Chromosoma. 1997 Jun;105(7-8):515–522. doi: 10.1007/BF02510488. [DOI] [PubMed] [Google Scholar]
  27. Morrissey J. P., Tollervey D. Yeast snR30 is a small nucleolar RNA required for 18S rRNA synthesis. Mol Cell Biol. 1993 Apr;13(4):2469–2477. doi: 10.1128/mcb.13.4.2469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mougey E. B., O'Reilly M., Osheim Y., Miller O. L., Jr, Beyer A., Sollner-Webb B. The terminal balls characteristic of eukaryotic rRNA transcription units in chromatin spreads are rRNA processing complexes. Genes Dev. 1993 Aug;7(8):1609–1619. doi: 10.1101/gad.7.8.1609. [DOI] [PubMed] [Google Scholar]
  29. Mougey E. B., Pape L. K., Sollner-Webb B. A U3 small nuclear ribonucleoprotein-requiring processing event in the 5' external transcribed spacer of Xenopus precursor rRNA. Mol Cell Biol. 1993 Oct;13(10):5990–5998. doi: 10.1128/mcb.13.10.5990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ni J., Tien A. L., Fournier M. J. Small nucleolar RNAs direct site-specific synthesis of pseudouridine in ribosomal RNA. Cell. 1997 May 16;89(4):565–573. doi: 10.1016/s0092-8674(00)80238-x. [DOI] [PubMed] [Google Scholar]
  31. Nicoloso M., Qu L. H., Michot B., Bachellerie J. P. Intron-encoded, antisense small nucleolar RNAs: the characterization of nine novel species points to their direct role as guides for the 2'-O-ribose methylation of rRNAs. J Mol Biol. 1996 Jul 12;260(2):178–195. doi: 10.1006/jmbi.1996.0391. [DOI] [PubMed] [Google Scholar]
  32. Pan Z. Q., Prives C. Assembly of functional U1 and U2 human-amphibian hybrid snRNPs in Xenopus laevis oocytes. Science. 1988 Sep 9;241(4871):1328–1331. doi: 10.1126/science.2970672. [DOI] [PubMed] [Google Scholar]
  33. Peculis B. A., Mount S. M. Ribosomal RNA: small nucleolar RNAs make their mark. Curr Biol. 1996 Nov 1;6(11):1413–1415. doi: 10.1016/s0960-9822(96)00745-2. [DOI] [PubMed] [Google Scholar]
  34. Peculis B. A., Steitz J. A. Disruption of U8 nucleolar snRNA inhibits 5.8S and 28S rRNA processing in the Xenopus oocyte. Cell. 1993 Jun 18;73(6):1233–1245. doi: 10.1016/0092-8674(93)90651-6. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Samarsky D. A., Schneider G. S., Fournier M. J. An essential domain in Saccharomyces cerevisiae U14 snoRNA is absent in vertebrates, but conserved in other yeasts. Nucleic Acids Res. 1996 Jun 1;24(11):2059–2066. doi: 10.1093/nar/24.11.2059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Savino R., Gerbi S. A. In vivo disruption of Xenopus U3 snRNA affects ribosomal RNA processing. EMBO J. 1990 Jul;9(7):2299–2308. doi: 10.1002/j.1460-2075.1990.tb07401.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schmitt M. E., Clayton D. A. Nuclear RNase MRP is required for correct processing of pre-5.8S rRNA in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Dec;13(12):7935–7941. doi: 10.1128/mcb.13.12.7935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shuai K., Warner J. R. A temperature sensitive mutant of Saccharomyces cerevisiae defective in pre-rRNA processing. Nucleic Acids Res. 1991 Sep 25;19(18):5059–5064. doi: 10.1093/nar/19.18.5059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Smith C. M., Steitz J. A. Sno storm in the nucleolus: new roles for myriad small RNPs. Cell. 1997 May 30;89(5):669–672. doi: 10.1016/s0092-8674(00)80247-0. [DOI] [PubMed] [Google Scholar]
  41. Tollervey D. Small nucleolar RNAs guide ribosomal RNA methylation. Science. 1996 Aug 23;273(5278):1056–1057. doi: 10.1126/science.273.5278.1056. [DOI] [PubMed] [Google Scholar]
  42. Trapman J., Planta R. J. Maturation of ribosomes in yeast. I Kinetic analysis by labelling of high molecular weight rRNA species. Biochim Biophys Acta. 1976 Sep 6;442(3):265–274. doi: 10.1016/0005-2787(76)90301-4. [DOI] [PubMed] [Google Scholar]
  43. Tycowski K. T., Shu M. D., Steitz J. A. Requirement for intron-encoded U22 small nucleolar RNA in 18S ribosomal RNA maturation. Science. 1994 Dec 2;266(5190):1558–1561. doi: 10.1126/science.7985025. [DOI] [PubMed] [Google Scholar]
  44. Tycowski K. T., Smith C. M., Shu M. D., Steitz J. A. A small nucleolar RNA requirement for site-specific ribose methylation of rRNA in Xenopus. Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14480–14485. doi: 10.1073/pnas.93.25.14480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Udem S. A., Warner J. R. Ribosomal RNA synthesis in Saccharomyces cerevisiae. J Mol Biol. 1972 Mar 28;65(2):227–242. doi: 10.1016/0022-2836(72)90279-3. [DOI] [PubMed] [Google Scholar]
  46. Venema J., Tollervey D. Processing of pre-ribosomal RNA in Saccharomyces cerevisiae. Yeast. 1995 Dec;11(16):1629–1650. doi: 10.1002/yea.320111607. [DOI] [PubMed] [Google Scholar]
  47. Warner J. R., Girard M., Latham H., Darnell J. E. Ribosome formation in HeLa cells in the absence of protein synthesis. J Mol Biol. 1966 Aug;19(2):373–382. doi: 10.1016/s0022-2836(66)80011-6. [DOI] [PubMed] [Google Scholar]
  48. Watkins N. J., Leverette R. D., Xia L., Andrews M. T., Maxwell E. S. Elements essential for processing intronic U14 snoRNA are located at the termini of the mature snoRNA sequence and include conserved nucleotide boxes C and D. RNA. 1996 Feb;2(2):118–133. [PMC free article] [PubMed] [Google Scholar]
  49. Xia L., Liu J., Sage C., Trexler E. B., Andrews M. T., Maxwell E. S. Intronic U14 snoRNAs of Xenopus laevis are located in two different parent genes and can be processed from their introns during early oogenesis. Nucleic Acids Res. 1995 Dec 11;23(23):4844–4849. doi: 10.1093/nar/23.23.4844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Xia L., Watkins N. J., Maxwell E. S. Identification of specific nucleotide sequences and structural elements required for intronic U14 snoRNA processing. RNA. 1997 Jan;3(1):17–26. [PMC free article] [PubMed] [Google Scholar]
  51. Yu Y. T., Shu M. D., Steitz J. A. A new method for detecting sites of 2'-O-methylation in RNA molecules. RNA. 1997 Mar;3(3):324–331. [PMC free article] [PubMed] [Google Scholar]
  52. Zafarullah M., Wisniewski J., Shworak N. W., Schieman S., Misra S., Gedamu L. Molecular cloning and characterization of a constitutively expressed heat-shock-cognate hsc71 gene from rainbow trout. Eur J Biochem. 1992 Mar 1;204(2):893–900. doi: 10.1111/j.1432-1033.1992.tb16709.x. [DOI] [PubMed] [Google Scholar]
  53. Zagorski J., Tollervey D., Fournier M. J. Characterization of an SNR gene locus in Saccharomyces cerevisiae that specifies both dispensible and essential small nuclear RNAs. Mol Cell Biol. 1988 Aug;8(8):3282–3290. doi: 10.1128/mcb.8.8.3282. [DOI] [PMC free article] [PubMed] [Google Scholar]

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