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Ultrastructural localization of transcription sites, DNA, and RNA reveals a concentric arrangement of structural and functional domains in plant nucleolonema

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Summary

By light microscopy the filamentous structure called nucleolonema was visualized in the nucleolus ofAllium cepa. Sites of rDNA transcription were located in the nucleolonema at the electron microscopic level. Thin sections of root tips were treated for desired times with bromo-UTP (BrUTP) and the sites of BrUTP incorporation were determined by immunogold labeling with antibromodeoxyuridine antibodies. The label was mostly seen at the periphery of the fibrillar centers and in the dense fibrillar component immediately surrounding them. This was principally the same in the specimens treated for 5, 10, and 30 min with BrUTP.The dense chromatin usually resided in the middle of the fibrillar centers and was surrounded by a light-fibrous material. An RNase-gold complex did not show any affinity to the fibrillar centers except their periphery. A combination of immunogold labeling for BrUTP incorporation and for DNA localization indicates that the rDNA transcription takes place in the circumferential zone around the dense chromatin. Immunogold labeling of BrUTP incorporation combined with osmium ammine-B staining for DNA suggests that the region around the dense chromatin containing a low concentration of DNA overlapped the site of rDNA transcription. The present data suggest that the transversal structural organization of the nucleolonema consists of the concentric arrangement of at least four different domains: the dense chromatin, the light-fibrous material intermingled with decondensed chromatin, rDNA transcription zone, and putative processing zone of rRNA transcripts.

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Abbreviations

DFC:

dense fibrillar component

FCs:

fibrillar centers

GC:

granular component

References

  • Bendayan M (1981) Ultrastructural localization of nucleic acids by the use of enzyme-gold complexes. J Histochem Cytochem 29: 531–541

    PubMed  Google Scholar 

  • Biggiogera M, Kaufmann SH, Shaper JH, Gas N, Amalric F, Fakan S (1991) Distribution of nucleolar proteins B23 and nucleolin during mouse spermatogenesis. Chromosoma 100: 162–172

    PubMed  Google Scholar 

  • Chouinard LA (1970) Localization of intranucleolar DNA in root meristematic cells ofAllium cepa. J Cell Sci 6: 73–85

    PubMed  Google Scholar 

  • De Cáreer G, Medina FJ (1999) Simultaneous localization of transcription and early processing markers allows dissection of functional domains in the plant cell nucleolus. J Struct Biol 128: 139–151

    PubMed  Google Scholar 

  • —, Cerdido A, Medina FJ (1997) NopA64, a novel nucleolar phosphoprotein from proliferating onion cells, sharing immunological determinants with mammalian nucleolin. Planta 201: 487–495

    PubMed  Google Scholar 

  • Dundr M, Raska I (1993) Nonisotopic ultrastructural mapping of transcription sites within the nucleolus. Exp Cell Res 208: 275–281

    PubMed  Google Scholar 

  • Estable C, Sotelo JR (1955) The behavior of the nucleolonema during mitosis. In: Symposium on fine structure of cells. Noordhoff, Groningen, pp 170–190

    Google Scholar 

  • Fakan S, Hernadez-Verdun D (1986) The nucleolus and the nucleolar organizer regions. Biol Cell 56: 189–205

    PubMed  Google Scholar 

  • Frens G (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nat Phys Sci 241: 20–22

    Google Scholar 

  • Goessens G, Lepoint A (1979) The nucleolus-organizing regions (NOR's): recent data and hypotheses. Biol Cell 35: 211–220

    Google Scholar 

  • Highett MI, Rawlins DJ, Shaw PJ (1993) Different patterns of rDNA distribution inPisum sativum nucleoli correlate with different levels of nucleolar activity. J Cell Sci 104: 843–852

    Google Scholar 

  • Hozák P (1995) Catching RNA polymerase I in flagranti: ribosomal genes are transcribed in the dense fibrillar component of the nucleolus. Exp Cell Res 216: 285–289

    PubMed  Google Scholar 

  • —, Cook PR, Schöfer C, Mosgöller W, Wachtier F (1994) Site of transcription of ribosomal RNA and intranucleolar structure in HeLa cells. J Cell Sci 107: 639–648

    PubMed  Google Scholar 

  • Hyde BB (1967) Changes in nucleolar ultrastructure associated with differentiation in the root tip. J Ultrastruct Res 18: 25–54

    PubMed  Google Scholar 

  • Iborra FJ, Pombo A, Jackson DA, Cook PR (1996) Active RNA polymerases are localized within discrete transcription “factories” in human nuclei. J Cell Sci 109: 1427–1436

    PubMed  Google Scholar 

  • Jackson DA, Hassan AB, Errington RJ, Cook PR (1993) Visualization of focal sites of transcription within human nuclei. EMBO J 12: 1059–1065

    PubMed  Google Scholar 

  • Jiménez-García LF, Segura-Valdez M de L, Ochs RL, Echeverria OM, Vázquez-Nin GH, Busch H (1993) Electron microscopic localization of ribosomal DNA in rat liver nucleoli by nonisotopic in situ hybridization. Exp Cell Res 207: 220–225

    PubMed  Google Scholar 

  • Jordan EG, Chapman JM (1971) Nucleolar and nuclear envelope ultrastructure in relation to cell activity in discs of carrot root (Dauern carota L). J Exp Bot 24: 197–209

    Google Scholar 

  • —, Bennett MD, Smith JB (1982) Nucleolar organizers and fibrillar centres inTriticum aestivum L., cv. Chinese Spring. Chromosoma 87: 447–459

    Google Scholar 

  • La Cour LF (1966) The internal structure of nucleoli. Chromosomes Today 1: 150–160

    Google Scholar 

  • Lafontaine JG, Lord A (1973) An ultrastructural and radioautographic investigation of the nucleolonemal component of plant interphase nucleoli. J Cell Sci 12: 369–383

    PubMed  Google Scholar 

  • Larsson M, Brundell E, Jörgensen P-M, Stahl S, Höög C (1999) Characterization of a novel nucleolar protein that transiently associates with the condensed chromosomes in mitotic cells. Eur J Cell Biol 78: 382–390

    PubMed  Google Scholar 

  • Lepoint A, Goessens G (1978) Nucleologenesis in Ehrlich tumour cells. Exp Cell Res 117: 89–94

    PubMed  Google Scholar 

  • Martin M, Moreno Díaz de la Espina S, Medina FJ (1989) Immunolocalization of DNA at nucleolar structural components in onion cells. Chromosoma 98: 368–377

    PubMed  Google Scholar 

  • Medina FJ, Risueño MC, Sánchez-Pina MA, Fernández-Gómez ME (1983) A study on nucleolar silver staining in plant cells: the role of argyrophilic proteins in nucleolar physiology. Chromosoma 88: 149–155

    Google Scholar 

  • Melcák I, Risueño MC, Raska I (1996) Ultrastructural nonisotopic mapping of nucleolar transcription sites in onion protoplasts. J Struct Biol 116: 253–263

    PubMed  Google Scholar 

  • Mineur P, Jennane A, Thiry M, Deltour R, Goessens G (1998) Ultrastructural distribution of DNA within plant meristematic cell nucleoli during activation and the subsequent inactivation by a cold stress. J Struct Biol 123: 199–210

    PubMed  Google Scholar 

  • Mosgöller W, Schöfer C, Derenzini M, Steiner M, Maier U, Wachtier F (1993) Distribution of DNA in human Sertoli cell nucleoli. J Histochem Cytochem 41: 1487–1493

    PubMed  Google Scholar 

  • Motte P, Deltour R, Mosen H, Bronchart R (1988) Threedimensional electron microscopy of the nucleolus and nucleolus-associated chromatin (NAC) during early germination ofZea mays L. Biol Cell 62: 65–81

    Google Scholar 

  • Olins AL, Moyer BA, Kim SH, Allison DP (1989) Synthesis of a more stable osmium ammine electron-dense DNA stain. J Histochem Cytochem 37: 395–398

    PubMed  Google Scholar 

  • Puvion-Dutilleul F, Bachellerie JP, Puvion E (1991) Nucleolar organization of HeLa cells as studied by in situ hybridization. Chromosoma 100: 395–409

    PubMed  Google Scholar 

  • Rawlins DJ, Shaw PJ (1990) Three-dimensional organization of ribosomal DNA in interphase nuclei ofPisum sativum by in situ hybridization and optical tomography. Chromosoma 99: 143–151

    Google Scholar 

  • Risueño MC, Medina FJ (1986) The nucleolar structure in plant cells. Cell Biol Rev 7: 1–140

    Google Scholar 

  • —, Testillano PS (1994) Cytochemistry and immunocytochemistry of nucleolar chromatin in plants. Micron 25: 331–360

    PubMed  Google Scholar 

  • —, Medina FJ, Moreno Díaz de la Espina (1982) Nucleolar fibrillar centres in plant meristematic cells: ultrastructure, cytochemistry and autoradiography. J Cell Sci 58: 313–329

    PubMed  Google Scholar 

  • Sato S (1988) Incorporation of argyrophilic proteins into the nucleolus-associated chromatin caused by exposure to actinomycin D. Biol Cell 63: 19–26

    PubMed  Google Scholar 

  • — (1992) Three-dimensional architecture of the higher plant nucleolonema disclosed on serial ultrathin sections. Biol Cell 75: 225–233

    Google Scholar 

  • —, Fujie T (1997) Architecture of the nucleolonema in root-tip cells ofRaphanus sativus. Cytologia 62: 411–419

    Google Scholar 

  • —, Yamada M (1996a) Effect of hypoxia on nucleoli in excised root tips ofVicia faba. Cytologia 61: 209–214

    Google Scholar 

  • Sato S, Yamada M (1996b) Effect of hypoxia on nucleoli in excised root tips ofVicia faba: an electron microscopic study. Cytologia 61: 269–276

    Google Scholar 

  • —, Yano H (1994) The nucleolonema changes its three-dimensional conformation during interphase in root-tip meristems of onion. Protoplasma 179: 172–180

    Google Scholar 

  • Scheer U, Raska I (1987) Immunocytochemical localization of RNA polymerase I in the fibrillar centers of nucleoli. Chromosomes Today 9: 284–293

    Google Scholar 

  • Shaw PJ, Jordan EG (1995) The nucleolus. Annu Rev Cell Biol 11: 93–121

    Google Scholar 

  • —, Highett MI, Beven AE Jordan EG (1995) The nucleolar architecture of polymerase I transciption and processing. EMBO J 14: 2896–2906

    PubMed  Google Scholar 

  • Shermoen AW, O'Farrell PH (1991) Progression of the cell cycle through mitosis leads to abortion of nascent transcripts. Cell 67: 303–310

    PubMed  Google Scholar 

  • Smetana K, Busch H (1974) The nucleolus and nucleolar DNA. In: Busch H (ed) The cell nucleus, vol 1. Academic Press, New York, pp 75–147

    Google Scholar 

  • Stahl A, Wachtier F, Hartung M, Devictor M, Schöfer C, Mosgöller W, De Lanversin A, Fouet C, Schwarzacher HG (1991) Nucleoli, nucleolar chromosomes and ribosomal genes in the human spermatocyte. Chromosoma 101: 231–244

    PubMed  Google Scholar 

  • Thiry M, Thiry-Blaise L (1989) In situ hybridization at the electron microscope level: an improved method for precise localization of ribosomal DNA and RNA. Eur J Cell Biol 50: 235–243

    PubMed  Google Scholar 

  • —, Scheer U, Goessens G (1988) Immunoelectron microscopic study of nucleolar DNA during mitosis in Ehrlich tumor cells. Eur J Cell Biol 47: 346–357

    PubMed  Google Scholar 

  • Thompson WF, Beven AF, Wells B, Shaw PJ (1997) Sites of rDNA transcription are widely dispersed through the nucleolus inPisum sativum and can comprise single genes. Plant J 12: 571–581

    PubMed  Google Scholar 

  • Wachtier F, Mosgöller W, Schwarzacher HG (1990) Electron microscopic in situ hybridization and autoradiography: localization and transcription of rDNA in human lymphocyte nucleoli. Exp Cell Res 187: 346–348

    PubMed  Google Scholar 

  • —, Schöfer C, Mosgöller W, Weipoltshammer K, Schwarzacher HG, Guichaoua M, Härtung M, Stahl A, Bergé-Lefranc JL, Gonzalez I, Sylvester J (1992) Human ribosomal RNA gene repeats are localized in the dense fibrillar component of nucleoli: light and electron microscopic in situ hybridization in human sertoli cells. Exp Cell Res 198: 135–143

    PubMed  Google Scholar 

  • Wansink DG, Nelissen RL, de Jong L (1994) In vitro splicing of pre-mRNA containing bromouridine. Mol Biol Rep 19: 109–113

    PubMed  Google Scholar 

  • Yano H, Sato S (1995) Cytochemical localization of intranucleolar DNA and RNA inAllium cepa root-tip cells. J Electron Microsc 44: 337–343

    Google Scholar 

  • — — (1999) Alterations of the intranucleolar DNA localization caused by hypoxic conditions in excised root tips ofAllium cepa. J Electron Microsc 48: 947–955

    Google Scholar 

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  1. Department of Biology and Earth Sciences, Faculty of Science, Ehime University, Bunkyo-cho 2-5, 790-8577, Matsuyama, Japan

    H. Yano & S. Sato

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  1. H. Yano
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  2. S. Sato
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Yano, H., Sato, S. Ultrastructural localization of transcription sites, DNA, and RNA reveals a concentric arrangement of structural and functional domains in plant nucleolonema. Protoplasma 214, 129–140 (2000). https://doi.org/10.1007/BF01279058

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  • DOI: https://doi.org/10.1007/BF01279058

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