Abstract
This work deals with the types of nuclear skeletal structures obtained from human fibroblast nuclei isolated by different procedures. It is confirmed that, in somatic vertebrate cells, the pore complex-lamina is always observed, whereas the presence of internal nucleolar and extranucleolar residual structures depends upon the method of nuclear isolation used. Furthermore, the results reported here argue for the existence of a nucleolar skeleton different from the nucleolar matrix often observed in different cell types by other investigators. The conditions of nuclear isolation which allow us to visualize this nucleolar skeleton without any other internal residual structures are described. The attachment of the nucleolar skeleton to the lamina suggested by the present data is considered in relation to the in situ position of nucleoli near the nuclear envelope.
Similar content being viewed by others
References
Aaronson RP, Blobel G (1975) Isolation of nuclear pore complexes in association with a lamina. Proc Natl Acad Sci USA 72:1007–1011
Adolph KW (1980) Organization of chromosomes in HeLa cells: Isolation of histone-depleted nuclei and nuclear scaffolds. J Cell Sci 42:291–304
Benavente R, Khrohne G, Schmidt-Zachmann MS, Hugle B, Franke WN (1984) Karyoskeletal proteins and the organization of the amphibian oocyte nucleus. J Cell Sci Suppl 1:161–186
Berezney R (1984) Organization and functions of the nuclear matrix. In: Hnilica LS (ed) Chromosomal non histone proteins. CRC Press, pp 119–180
Berezney R, Coffey DS (1974) Identification of a nuclear protein matrix. Biochem Biophys Res Commun 60:1410–1417
Berezney R, Coffey DS (1977) Nuclear matrix. Isolation and characterization of a framework structure from rat liver nuclei. J Cell Biol 73:616–637
Bolla RS, Braaten DC, Shiomi Y, Hebert MB, Schlessinger D (1985) Localization of specific rDNA spacer sequences to the mouse L-cell nucleolar matrix. Mol Cell Biol 5:1287–1294
Bourgeois CA, Hemon D, Bouteille M (1979) Structural relationship between the nucleolus and the nuclear envelope. J Ultrastruct Res 68:328–340
Bourgeois CA, Laquerrière F, Hémon D, Hubert J, Bouteille M (1985) New data on the in situ position of the inactive X chromosome in the interphase nucleus of human fibroblasts. Hum Genet 69:122–129
Bouteille M, Bouvier D, Sève AP (1983) Heterogeneity and territorial organization of the nuclear matrix and related structures. Int Rev Cytol 83:135–182
Bouvier D, Hubert J, Bouteille M (1980) The nuclear shell in HeLa cell nuclei: Whole mount electron microscopy of the dissociated and isolated nuclear periphery. J Ultrastruct Res 73:288–298
Bouvier D, Hubert J, Sève AP, Bouteille M (1982) RNA is responsible for three-dimensional organization of nuclear matrix proteins in HeLa cells. Biol Cell 43:143–146
Bouvier D, Hubert J, Sève AP, Bouteille M (1985a) Nuclear RNAassociated proteins and their relationship to the nuclear matrix and related structures in HeLa cells. Can J Biochem Cell Biol 63:631–643
Bouvier D, Hubert J, Sève AP, Bouteille M (1985b) Characterization of lamina-bound chromatin in the nuclear shell isolated from HeLa cells. Exp Cell Res 156:500–512
Bouvier D, Hubert J, Sève AP, Bouteille M (1985c) Structural aspects of intranuclear matrix disintegration upon RNase digestion of HeLa cell nuclei. Eur J Cell Biol 36:323–333
Bureau J, Hubert J, Bouteille M (1986) Attachment of DNA to nucleolar and nuclear skeletal structures as visualized by Kleinschmidt molecular spreading. Biol Cell 56:61–70
Daskal Y, Busch H (1978) Ultrastructure of chromatin and chromosomes as visualized by scanning electron microscopy. In: Busch H (ed) The cell nucleus, vol IV. Academic Press, New York, pp 3–45
Gautier A (1976) Ultrastructural localization of DNA in ultrathin tissue sections. Int Rev Cytol 44:113–191
Gerace L, Blobel G (1982) Nuclear lamina and the structural organization of the nuclear envelope. Cold Spring Harbor Symp Quant Biol 46:967–978
Gerace L, Blum A, Blobel G (1978) Immunocytochemical localization of the major polypeptide of the nuclear pore complex lamina fraction. J Cell Biol 79:546–566
Gerace L, Conneau C, Benson M (1984) Organization and modulation of nuclear lamina structure. J Cell Sci Suppl 1:137–160
Goessens G (1984) Nucleolar structure. Int Rev Cytol 87:107–158
Granick D (1975) Nucleolar necklaces in chick embryo fibroblast cells. II Microscope observation of the effect of adenosine analogues on nucleolar necklace formation. J Cell Biol 65:418–427
Hancock R (1974) Interphase chromosomal deoxyribonucleoproteins isolated as a discrete structure from cultured cells. J Mol Biol 86:649–663
Hancock R (1982) Topological organisation of interphase DNA: the nuclear matrix and other skeletal structures. Biol Cell 46:105–122
Hancock R, Hughes ME (1982) Organisation of DNA in the interphase nucleus. Biol Cell 44:201–212
Hubert J, Bourgeois CA (1986) The nuclear skeleton and the spatial arrangement of chromosomes in the interphase nucleus of vertebrate somatic cell. Hum Genet 74:1–15
Hubert J, Bouvier D, Bouteille M (1979) Nuclear cortex: Dissociation of nuclei isolated from cultured human cells. Biol Cell 36:87–90
Hubert J, Bureau J, Bouteille M (1984) Anchorage of the nucleolus in the pore complex-lamina by a DNA-bearing structure masked in situ in rat liver nuclei. Biol Cell 52:91–102
Kaufmann SH, Shaper JH (1984) A subset of non histone nuclear proteins reversibly stabilized by the sulfydryl cross-linking reagent tetrathionate. Exp Cell Res 155:477–495
Kaufmann SH, Coffey DS, Shaper JH (1981) Considerations in the isolation of rat liver nuclear matrix, nuclear envelope and pore complex-lamina. Exp Cell Res 132:105–123
Kaufmann SH, Gibson W, Shaper JH (1983) Characterization of the major polypeptides of rat liver nuclear envelope. J Biol Chem 258:2710–2719
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lebkowski JS, Laemmli UK (1982a) Evidence for two levels of DNA folding in histone-depleted HeLa interphase nuclei. J Mol Biol 156:309–324
Lebkowski JS, Laemmli UK (1982b) Non-histone proteins and long-range organization of HeLa interphase DNA. J Mol Biol 156:325–344
Lewis CD, Lebkowski JS, Daly AK, Laemmli UK (1984) Interphase nuclear matrix and metaphase scaffolding structures. J Cell Sci Suppl 1:103–122
Long BH, Ochs RL (1983) Nuclear matrix, hnRNA and snRNA in Friend erythroleukemia nuclei depleted of chromatin by low ionic strength EDTA. Biol Cell 48:89–98
Miller TE, Huang CY, Pogo AO (1978) Rat liver nuclear skeleton and ribonucleoprotein complexes containing HnRNA. J Cell Biol 76:675–691
Rae PMM, Franke WW (1972) The interphase distribution of satellite DNA-containing heterochromatin. Chromosoma 39:443–456
Roels F, Goldfischer S (1971) Staining of nucleic acids with 3,3′- diaminobenzidine (DAB). J Histochem Cytochem 19:713–714
Smetana K, Daskal I, Busch H (1980) Cytochemistry of the microtrabecular network in compact nucleoli of hepatocytes treated with cycloheximide. Histochemistry 65:301–308
Van Eekelen CAG, Salden MHL, Habets WJA, Van de Putte LBA, Van Venrooij WJ (1982) On the existence of an internal nuclear protein structure in HeLa cells. Exp Cell Res 141:181–190
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bourgeois, C.A., Bouvier, D., Seve, A.P. et al. Evidence for the existence of a nucleolar skeleton attached to the pore complex-lamina in human fibroblasts. Chromosoma 95, 315–323 (1987). https://doi.org/10.1007/BF00293178
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00293178