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Localization of snRNP antigens in nucleolus-associated bodies: study of plant interphase nuclei by confocal and electron microscopy

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Abstract

Using two monoclonal antibodies, mAb Y12 and mAbK121, small nuclear ribonucleoproteins (snRNPs) were localized by immunofluorescence and immunogold electron microscopy in Pisum sativum and Cicer arietinum interphase nuclei. We showed that snRNPs were mostly found in nucleolus associated bodies (NABs) characterizing these two plant species. snRNPs were also found threoughout the nucleoplasm, their distribution being diffuse or speckled depending on the nucleus. Examination of optical sectins furnished by confocal laser microscopy allowed us to obtain more precise information on the number and location of NABs. One, two and rarely three (in green pea) of such structures were observed, their size varied from 0.8 to 1.5 μm and they were also systematically observed in intimate contact with the nucleolus. Under electron microscopy, labelling was likewise found to be noticeably higher over NABs than over the nucleoplasm. The possible relationship between NABs and other nuclear bodies found in animal cells and known to be involved in splicing of pre-mRNA is discussed.

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References

  • Barlow PW (1983a) Nucleolus-associated bodies (karyosomes) in dividing and differentiating plant cells. Protoplasma 115:1–10

    Google Scholar 

  • Barlow PW (1983b) Changes in the frequency of two types of nuclear body during the interphase of meristematic plant cells. Protoplasma 118:104–113

    Google Scholar 

  • Bennett MD, Smith JB, Ward J, Jenkins G (1980) The relationship between nuclear DNA content and centromere volume in higher plants. J Cell Sci 47:91–115

    Google Scholar 

  • Callan HG, Gall JG (1991) Association of RNA with the B and C snurposomes of Xenopus oocyte nuclei. Chromosoma 101:69–82

    Google Scholar 

  • Carmo-Fonseca M, Pepperkok R, Carvalho MT, Lamond AI (1992) Transcription-dependent colocalization of the U1, U2, U4/U6 and U5 snRNPs in coiled bodies. J Cell Biol 117:1–14

    Google Scholar 

  • Chaly NM, Setterfield G (1975) Organization of the nucleus, nucleolus and protein-synthesizing apparatus in relation to cell development in roots of Pisum sativum. Can J Bot 53:200–218

    Google Scholar 

  • Chang TH, Clark MW, Lustig AJ, Cusick ME, Abelson J (1988) RNA 11 protein is associated with the yeast spliceosome and is localized in the periphery of the nucleus. Mol Cell Biol 8:2379–2393

    Google Scholar 

  • Church K, Moens P (1976) Centromere behavior during interphase and meiotic prophase in Allium fistulosum from 3-D, EM reconstruction. Chromosoma 56:249–263

    Google Scholar 

  • Egeland DB, Sturtevant AP, Schuler MA (1989) Molecular analysis of dicot and monocot small nuclear RNA populations. Plant Cell 1:633–643

    Google Scholar 

  • Fakan S, Leser G, Martin TE (1984) Ultrastructural distribution of nuclear ribonucleoproteins as visualized by immunocytochemistry on thin sections. J Cell Biol 98:358–363

    Google Scholar 

  • Gall JG (1991) Spliceosomes and snurposomes Science 252:1499–1500

    Google Scholar 

  • Gall JG, Callan HG (1989) The sphere organelle contains small ribonucleoproteins. Proc Natl Acad Sci USA 86:6635–6639

    Google Scholar 

  • Gillies CB (1973) Ultrastructural analysis of maize pachytene karyotype by three dimensional reconstruction of the synaptonemal complexes. Chromosoma 43:145–176

    Google Scholar 

  • Johnson DA, Gautsch JW, Sportsman JR, Elder JE (1984) Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Techniques 1:3–8

    Google Scholar 

  • Jordan EG (1976) Nuclear structure in Daucus carota L. Cytobiologie 14:171–177

    Google Scholar 

  • Kiss T, Tóth M, Solymosy F (1985) Plant small nuclear RNAs. Nucleolar U3 snRNA is present in plants: partial characterization. Eur J Biochem 152:259–266

    Google Scholar 

  • Kiss T, Antal M, Solymosy F (1987a) Plant small nuclear RNAs. II U6 RNA and a 4.5 S-like RNA are present in plant nuclei. Nucleic Acids Res 15:543–560

    Google Scholar 

  • Kiss T, Antal M, Solymosy MF (1987b) Plant small nuclear RNAs. III The complete primary and secondary structure of broad bean U2: Phylogenetic and functional implications. Nucleic Acids Res 15:1332

    Google Scholar 

  • Kiss T, Jakab G, Antal M, Palfi H, Kis M, Solymosy MF (1988) Plant small nuclear RNAs. V U4 RNA is present in broad bean plants in the form of sequence variants and is base-paired with U6 RNA. Nucleic Acids Res 16:5407–5426

    Google Scholar 

  • Krainer AR (1988) Pre-mRNA splicing by complementation with purified human U1, U2, U4/U6 snRNPs. Nucleic Acids Res 16:9415–9429

    Google Scholar 

  • Krol A, Ebel JP (1983) U1, U2 and U5 small nuclear RNAs are found in plant cells. Complete nucleotide sequence of the U5 RNA family from pea nuclei. Nucleic acids Res 11:8583–8594

    Google Scholar 

  • Lafontaine JG (1965) A light and electron microscope study of small, spherical nuclear bodies in meristematic cells of Allium cepa, Vicia faba, Raphanus sativus. J Cell Biol 26:1–17

    Google Scholar 

  • Lafontaine JG (1968) Structural components of the nucleus in mitotic plant cells. In: Dalton AJ, Haguenau F (eds) Ultrastructure in biological systems: The nucleus, vol 3. Academic Press, New York, pp 151–196

    Google Scholar 

  • Lafontaine JG, Luck BT (1980) An ultrastructural study of plant cells (Allium porrum) centromeres. J Ultrastruct Res 70:298–307

    Google Scholar 

  • Lafontaine JG, Luck BT, Gugg S (1991) Nucleolus-associated bodies in meristematic cells of two plants (Cicer arietinum, Leucaena glauca) with different ploidy levels. Can J Bot 69:1329–1336

    Google Scholar 

  • Lerner MR, Boyle JA, Mount SM, Wolin SL, Steitz JA (1980) Are snRNPs involved in splicing? Nature 283:220–224

    Google Scholar 

  • Luck BT, Lafontaine JG (1982) An ultracytochemical study of nuclear bodies in meristematic plant cells (Cicer arietinum). Can J Bot 60:611–619

    Google Scholar 

  • Maniatis T, Reed R (1987) The role of small nuclear ribonucleoprotein particles in pre-mRNA splicing. Nature 325:673–678

    Google Scholar 

  • Medina FJ, Risueño MC (1981) Nucleolar structure and dynamics during meiotic prophase in pea ovules. Biol Cell 42:79–86

    Google Scholar 

  • Raska I, Ochs RL, Andrade LEC, Chan EKL, Burlingame R, Peebles C, Gruol D, Tan EM (1990) Association between the nucleolus and the coiled body. J Struct Biol 104:120–127

    Google Scholar 

  • Raska I, Andrade LEC, Ochs RL, Chan EKL, Chang CM, Roos G, Tan EM (1991) Immunological and ultrastructural studies of the nuclear coiled body with autoimmune anribodies. Exp Cell Res 195:27–37

    Google Scholar 

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

    Google Scholar 

  • Reuter R, Appel B, Bringmann P, Rinke J, Lührmann R (1984) 5′-terminal caps of snRNAs are reactive with antibodies specific for 2,2,7-trimethylguanosine in whole cells and nuclear matrices. Exp Cell Res 154:548–460

    Google Scholar 

  • Simmonds DH, Seagull RW, Setterfield G (1985) Evaluation of techniques for immunofluorescent staining of microtubules in cultured plant cells. J Histochem Cytochem 33:345–352

    Google Scholar 

  • Vásquez-Nin GH, Echeverría OM, Mínguez A, Moreno Díaz de la Espina S, Fakan S, Martin TE (1992) Ribonucleoprotein components of root meristematic cell nuclei of the tomato characterized by application of mild loosening and immunocytochemistry. Exp Cell Res 200:431–438

    Google Scholar 

  • Weber K, Rathke PC, Osborne M (1978) Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cell by electron microscopy and by immunofluorescence microscopy. Proc Natl Acad Sci USA 75:1820–1824

    Google Scholar 

  • Williams LM, Jordan EG, Barlow PW (1983) The ultrastructure of nuclear bodies in interphase plant cell nuclei. Protoplasma 118:95–103

    Google Scholar 

  • Williams LM, Charest PM, Fitzgerald GJ, Lafontaine JG (1985) A comparison of nuclease-gold and protease-gold complex labeling over the nucleolus and nuclear bodies of Pisum sativum root tip cells. Biol Cell (Paris) 54:65–72

    Google Scholar 

  • Wu Z, Murphy C, Callan HG, Gall JG (1991) Small nuclear ribonucleoproteins in the amphibian germinal vesicle: loops, spheres, and snurposomes. J Cell Biol 113:465–483

    Google Scholar 

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Communicated by: J.B. Rattner

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Chamberland, H., Lafontaine, J.G. Localization of snRNP antigens in nucleolus-associated bodies: study of plant interphase nuclei by confocal and electron microscopy. Chromosoma 102, 220–226 (1993). https://doi.org/10.1007/BF00352395

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  • DOI: https://doi.org/10.1007/BF00352395

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