Abstract
Mature Balbiani Ring (BR) granules in situ were stained with the nucleic acid specific stain, osmium ammine-B, recorded by electron spectroscopic imaging and reconstructed by electron microscope tomography to examine the three-dimensional (3-D) distribution of BR heterogeneous nuclear RNA (hnRNA). The BR2 granules contain ca. 37 kb of mRNA. Reconstructed BR granules were selected to emphasize one of the prevalent conformations seen in the sectioned salivary glands, the en face or “pin-wheel” conformation. A variety of image processing and volume-rendering operations were applied to the set of reconstructed BR granules. Some of the conclusions of this study are the following: (1) RNA distribution is not uniform throughout the granule; (2) RNA is condensed into about ten particles per granule, which all appear to possess approximately the same RNA stain density; (3) heterogeneity exists in the positions and sizes of particles within the various BR granules. These data argue for the folding of a beaded ribbon, consisting of connected particulate condensations of BR mRNA, possessing considerable 3-D flexibility, even in the packaged state. A comparison of this beadedribbon model and a prior folded hnRNP fiber model is also presented.
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References
Baurén G, Wieslander L (1994) Splicing of Balbiani ring 1 gene pre-mRNA occurs simultaneously with transcription. Cell 76:183–192
Beermann W (1952) Chromomerenkonstanz und spezifische Modifikationen der Chromosomenstruktur in der Entwichlung und Organdifferenzierung von Chironomus tentans. Chromosoma 5:139–198
Beermann W, Bahr GF (1954) The submicroscopic structure of the Balbiani ring. Exp Cell Res 6:195–201
Case ST, Wieslander L (1992) Results Probl Cell Differ 19:187–226
Conway G, Wooley J, Bibring T, LeStourgeon WM (1988) Ribonucleoproteins package 700 nucleotides of pre-mRNA into a repeating array of regular particles. Mol Cell Biol 8:2884–2895
Daneholt B (1982) Structural and functional analysis of Balbiani ring genes in the salivary glands of Chironomus tentans. In: King RC, Akai H (eds) Insect ultrastructure. Plenum Press, New York, pp 382–401
Daneholt B, Björkroth B, Lönnroth A, Mehlin H, Sköglund U (1990) Synthesis and transport of a specific premessenger RNP particle. Chromosomes Today 10:90–100
Derenzini M, Farabegoli F (1990) Selective staining of nucleic acids by osmium-ammine complex in thin sections from lowicryl-embedded samples. J Histochem Cytochem 38:1495–1501
Dreyfuss G, Matunis MJ, Seraffin P-R, Burd CG (1993) hnRNP proteins and the biogenesis of mRNA. Annu Rev Biochem 62:289–321
Grond C, Saiga H, Edström J-E (1987) The sp-I genes in the Balbiani rings of Chironomus salivary glands. In: Hennig W (ed) Structure and function of eukaryotic chromosomes (Results and problems in cell differentiation 14). Springer, Berlin Heidelberg New York, pp 69–80
Hamming RW (1983) Digital filters. Prentice-Hall, Englewood Cliffs, NY, p 257
Keene JD, Query CC (1991) Nuclear RNA-binding proteins. Prog Nucleic Acid Res 41:179–202
Lamvik MK, Davilla SD (1988) Irradiation-induced mass loss effects at low temperature are dependent on specimen material. In: Bailey GW (ed) Proceedings of the 46th Annual Meeting of the Electron Microscopy Society of America. San Francisco Press, California, pp 636–637
Lönnroth A, Alexciev K, Mehlin H, Wurtz T, Sköglund U, Daneholt B (1992a) Demonstration of a 7-nm RNP fiber as a basic structural element in a premessenger RNP particle. Exp Cell Res 199:292–296
Lönnroth A, Sköglund U, Tokuyasu KT, Daneholt B (1992b) The adsorption staining technique applied to isolated premessenger ribonucleoprotein particles: a comparison with conventional techniques using electron microscope tomography. J Microsc. 170:173–182
Luther PK, Lawrencre MC, Crowther RA (1988) A method for monitoring the collapse of plastic sections as a function of electron dose. Ultramicroscopy 24:7–18
Mehlin H, Lönnroth A, Sköglund U, Daneholt B (1988) Structure and transport of a specific premessenger RNP particle. Cell Biol Int Rep 12:729–736
Mehlin H, Sköglund U, Daneholt B (1991) Transport of Balbiani ring granules through nuclear pores in Chironomus tentans. Exp Cell Res 193:72–77
Merrill BM, Williams KR (1990) Structure/function relationships in hnRNP proteins. In: Straus PR, Wilson SH (eds) The eukaryotic nucleus: molecular biochemistry and macromolecular assemblies, vol 2. The Telford Press, Caldwell, New Jersey, pp 579–604
Meyer B, Mähr R, Eppenberger HM, Lezzi M (1983) The activity of the Balbiani rings 1 and 2 in salivary glands of Chironomus tentans under different modes of development and after pilocarpine treatment. Dev Biol 98:265–277
Olins AL, Olins DE, Franke WW (1980) Stereo-electron microscopy of nucleoli, Balbiani rings and endoplasmic reticulum in Chironomus salivary gland cells. Eur J Cell Biol 22:714–723
Olins AL, Moyer BA, Kim S-H, Allison DP (1989) Synthesis of a more stable osmium ammine electron-dense DNA stain. J Histochem Cytochem 37:395–398
Olins AL, Olins DE, Bazett-Jones DP (1992) Balbiani ring hnRNP substructure visualized by selective staining and electron spectroscopic imaging. J Cell Biol 117:483–491
Olins AL, Olins DE, Levy HA, Shah MB, Bazett-Jones DP (1993) Electron microscope tomography of Balbiani ring hnRNP substructure. Chromosoma 102:137–144
Olins DE, Olins AL, Levy HA, Durfee RC, Margle SM, Tinnel EP, Dover SD (1983) Electron microscope tomography: transcription in three dimensions. Science 220:498–500
Osheim YN, Beyer AL (1991) EM analysis of Drosophila chorion genes: amplification, transcription termination and RNA splicing. Electron Microsc Rev 4:111–128
Paulsson G, Höög C, Bernholm K, Wieslander L (1992) Balbiani ring 1 gene in Chironomus tentans sequence organization and dynamics of a coding minisatellite. J Mol Biol 225:349–361
Pelling C (1964) Ribonukleinsaure-Synthese der Riesenchromosomen. Autoradiographische Untersuchungen an Chironomus tentans. Chromosoma 15:71–122
Rech JE, Huang J, LeStourgeon WM (1994) The packaging of premessenger RNA. In: Krainer A (ed) RNA Processing. Oxford Univ Press (in press)
Sköglund M, Anderson K, Björkroth B, Lamb MM, Daneholt B (1983) Visualization of the formation and transport of a specific hnRNP particle. Cell 34:847–855
Sköglund U, Daneholt B (1986) Electron microscope tomography. Trends Biochem Sci 11:499–503
Sköglund U, Anderson K, Strandberg B, Daneholt B (1986) Three-dimensional structure of a specific premessenger RNP established by electron microscope tomography. Nature 319:560–564
Stevens BJ, Swift H (1966) RNA transport from nucleus to cytoplasm in Chironomus salivary glands. J Cell Biol 31:55–77
Trepte H-H (1993) Ultrastructural analysis of Balbiani ring genes of Chironomus pallidivittatus in different states of Balbiani ring activity. Chromosoma 102:403–445
Vazquez-Nin GH, Bernhard W (1971) Comparative ultrastructural study of perichromatin and Balbiani ring granules. J Ultrastruct Res 36:842–860
Wieslander L, Paulsson G (1992) Sequence organization of the Balbiani ring 2.1 gene in Chironomus tentans. Proc Natl Acad Sci USA 89:4578–4582
Wurtz T, Lönnroth A, Ovchinnikov L, Sköglund U, Daneholt B (1990a) Isolation and initial characterization of a specific premessenger ribonucleoprotein particle. Proc Natl acad Sci USA 87:831–835
Wurtz T, Lönnroth A, Daneholt B (1990b) Higher order structure of Balbiani ring premessenger RNP particles depends on certain RNAse A sensitive sites. J Mol Biol 215:93–101
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Olins, A.L., Olins, D.E., Olman, V. et al. Modeling the 3-D RNA distribution in the Balbiani Ring granule. Chromosoma 103, 302–310 (1994). https://doi.org/10.1007/BF00417877
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DOI: https://doi.org/10.1007/BF00417877