Abstract
We investigated the presence and localization, in the cells of anucleolate mutant embryos ofXenopus laevis, of three representative small nucleolar RNAs (snoRNAs), U3, U15 and U17, and of two nucleolar proteins, nucleolin and fibrillarin. The levels of the three snoRNAs in the anucleolate mutant are the same as in normal embryos, in contrast to 5S RNA and ribosomal proteins. In situ hybridization showed that, in the absence of fully organized nucleoli, the three RNAs are diffusely distributed in the nucleus and partly associated with a number of small structures. Nucleolin and fibrillarin are also present in the anucleolate embryos as in normal embryos, although there is less nucleolin mRNA in the former. The two nucleolar proteins were localized by immunofluorescence microscopy. Fibrillarin, similar to its associated U3 and U15 snoRNAs, is diffusely distributed in the anucleolate nucleus and is partly associated with small structures, probably prenucleolar bodies and pseudonucleoli. Nucleolin also appears diffusely distributed in the nucleus with some spots of higher concentration, but with a different pattern with respect to fibrillarin.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Balakin AG, Smith L, Fournier MJ (1996) The RNA world of the nucleolus: two major families of small RNAs defined by different box elements with related functions. Cell 86:823–834
Baserga SJ, Yang XW, Steitz JA (1991) 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 10:2645–2651
Brown DD (1967) The genes for ribosomal RNA and their transcription during amphibian development. Curr Top Dev Biol 2:47–73
Brown DD, Gurdon JB (1964) Absence of ribosomal RNA synthesis in the anucleolate mutant ofXenopus laevis. Proc Natl Acad Sci USA 51:139–146
Brown DD, Weber CS (1968) Gene linkage by RNA-DNA hybridization. I. Unique DNA sequences homologous to 4S RNA, 5S RNA, and ribosomal RNA. J Mol Biol 34:661–680
Caizergues-Ferrer M, Mariottini P, Curie C, Lapeyre B, Gas N, Amalric F, Amaldi F (1989) Nucleolin fromXenopus laevis: cDNA cloning and expression during development. Genes Dev 3:324–333
Caizergues-Ferrer M, Mathieu C, Mariottini P, Amalric F, Amaldi F (1991) Developmental expression of fibrillarin and U3 snRNA inXenopus laevis. Development 112:317–326
Cecconi F, Mariottini P, Loreni F, Pierandrei-Amaldi P, Campioni N, Amaldi F (1994) U17XS8, a small nucleolar RNA with a 12 nt complementarity to 18S rRNA and coded by a sequence repeated in the six introns ofXenopus laevis r-protein S8 gene. Nucleic Acids Res 22:732–741
De La Torre C, Gimenez-Martin G (1982) The nucleolus cycle. In: Jordan EG, Cullis CA (eds) The nucleolus. Cambridge University Press, Cambridge, UK, pp 153–177
Elsdale TR, Fischberg M, Smith S (1958) A mutation that reduces nucleolar number inXenopus laevis. Exp Cell Res 14:642–643
Esper H, Barr HJ (1964) A study of the developmental cytology of a mutation affecting nucleoli inXenopus embryos. Dev Biol 10:105–121
Hadjiolov AA (1985) The nucleolus and ribosome biogenesis. Springer, Berlin
Hay ED, Gurdon JB (1967) Fine structure of the nucleolus in normal and mutantXenopus embryos. J Cell Sci 2:151–162
Hughes JMX, Ares M (1991) 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 10:4231–4239
Jeppesen C, Stebbins-Boaz B, Gerbi SA (1988) Nucleotide sequence determination and secondary structure ofXenopus U3 snRNA. Nucleic Acids Res 16:2127–2148
Kass F, Tyc K, Steitz JA, Sollner-Webb B (1990) The U3 small nucleolar ribonucleoprotein functions in the first step of preribosomal processing. Cell 60:897–908
Kiss-László Z, Henry Y, Bachellerie JP, Caizergues-Ferrer M, Kiss T (1996) Site-specific ribose methylation of preribosomal RNA: a novel function for small nucleolar RNAs. Cell 85:1077–1088
Lapeyre B, Mariottini P, Mathieu C, Ferrer P, Amaldi F, Amalric F, Caizergues-Ferrer M (1990) Molecular cloning ofXenopus fibrillarin, a conserved U3 small nuclear ribonucleoprotein recognized by antisera from humans with autoimmune disease. Mol Cell Biol 10:430–434
Maxwell ES, Fournier MJ (1995) The small nucleolar RNAs. Annu Rev Biochem 35:897–934
Messmer B, Dreyer C (1993) Requirements for nuclear translocation and nucleolar accumulation of nucleolin ofXenopus laevis. Eur J Cell Biol 61:369–382
Miller L (1973) Control of 5S RNA synthesis during early development of anucleolate and partial nucleolate mutants ofXenopus laevis. J Cell Biol 59:624–532
Miller L (1974) Metabolism of 5S RNA in the absence of ribosome production. Cell 3:275–281
Nicoloso M, Qu LH, Michot B, Bachellerie JP (1996) 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 260:178–195
Ochs RL, Lischwe MA, Shen E, Carroll RE, Busch H (1985) Nucleologenesis: composition and fate of prenucleolar bodies. Chromosoma 92:330–336
Olson MO (1990) The role of proteins in nucleolar structure and function. In: Strauss PR, Wilson SH (eds) The eukaryotic nucleus. Telford Press, Caldwell, NJ, pp 519–577
Pellizzoni L, Crosio C, Campioni N, Loreni F, Pierandrei-Amaldi P (1994) Different forms of U15 snoRNA are encoded in the introns of the ribosomal protein S1 gene ofXenopus laevis. Nucleic Acids Res 22:4607–4613
Pellizzoni L, Cardinali B, Lin-Marq N, Mercanti D, Pierandrei-Amaldi P (1996) AXenopus laevis homologue of the La autoantigen binds the pyrimidine tract of the 5'UTR of ribosomal protein mRNAs in vitro: implication of a protein factor in complex formation. J Mol Biol 259:904–915
Pieler T, Appel B, Oei SL, Mentrel H, Erdmann A (1985) Point mutational analysis of theXenopus laevis 5S gene promoter. EMBO J 4:1847–1853
Pierandrei-Amaldi P, Amaldi F (1994) Aspects of regulation of ribosomal protein synthesis inXenopus laevis. Review. Genetica 94:181–193
Pierandrei-Amaldi P, Campioni N, Beccari E, Bozzoni I (1982) Expression of ribosomal protein genes inXenopus laevis development. Cell 30:163–171
Pierandrei-Amaldi P, Beccari E, Bozzoni I, Amaldi F (1985) Ribosomal protein production in normal and anucleolateXenopus embryos: regulation at the posttranscriptional and translational levels. Cell 42:317–323
Probst EA, Kressman A, Birnstiel ML (1979) Expression of sea urchin histone genes in the oocyte ofXenopus laevis. J Mol Biol 135:709–732
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Savino R, Gerbi SA (1990) In vivo disruption ofXenopus U3 snRNA affects ribosomal RNA processing. EMBO J 9:2299–2308
Scheer U, Benavente R (1990) Functional and dynamic aspects of the mammalian nucleolus. BioEssays 12:14–21
Steele RE, Thomas PS, Reeder RH (1984) Anucleolate frog embryos contain ribosomal DNA sequences and a nucleolar antigen. Dev Biol 102:409–416
Tashiro K, Shiokawa K, Yamana K, Sakaki Y (1986) Structural analysis of ribosomal DNA homologues in nucleolus-less mutant ofXenopus laevis. Gene 44:299–306
Wallace H, Birnstiel ML (1966) Ribosomal cistrons and the nucleolar organizer. Biochim Biophys Acta 114:296–310
Wilkinson DG (1992) The theory and practice of in situ hybridization. In: Wilkinson DG (ed) In situ hybridization: a practical approach. IRL Press, Oxford, pp 1–13
Author information
Authors and Affiliations
Corresponding author
Additional information
Edited by: S.A. Gerbi
Rights and permissions
About this article
Cite this article
Crosio, C., Campioni, N., cardinali, B. et al. Small nucleolar RNAs and nucleolar proteins inXenopus anucleolate embryos. Chromosoma 105, 452–458 (1997). https://doi.org/10.1007/BF02510482
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02510482