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J Cell Biol. Nov 1, 1978; 79(2): 546–566.
PMCID: PMC2110258

Immunocytochemical localization of the major polypeptides of the nuclear pore complex-lamina fraction. Interphase and mitotic distribution

Abstract

This laboratory has previously isolated a fraction from rat liver nuclei consisting of nuclear pore complexes associated with the proteinaceous lamina which underlies the inner nuclear membrane. Using protein eluted from sodium dodecyl sulfate (SDS) gels, we have prepared antibodies in chickens to each of the three predominant pore complex- lamina bands. Ouchterlony double diffusion analysis shows that each of these individual bands cross-reacts strongly with all three antisera. In immunofluorescence localization performed on tissue culture cells with these antibodies, we obtain a pattern of intense staining at the periphery of the interphase nucleus, with little or no cytoplasmic reaction. Electron microscope immunoperoxidase staining of rat liver nuclei with these antibodies labels exclusively the nuclear periphery. Furthermore, reaction occurs in areas which contain the lamina, but not at the pore complexes. While our isolation procedure extracts the internal contents of nuclei completely, semiquantitative Ouchterlony analysis shows that it releases negligible amounts of these lamina antigens. Considered together, our results indicate that these three bands represent major components of a peripheral nuclear lamina, and are not structural elements of an internal "nuclear protein matrix." Fluorescence microscopy shows that the perinuclear interphase localization of these lamina proteins undergoes dramatic changes during mitosis. Concomitant with nuclear envelope disassembly in prophase, these antigens assume a diffuse localization throughout the cell. This distribution persists until telophase, when the antigens become progressively and completely localized at the surface of the daughter chromosome masses. We propose that the lamina is a biological polymer which can undergo reversible disassembly during mitosis.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Adolphs KW, Cheng SM, Paulson JR, Laemmli UK. Isolation of a protein scaffold from mitotic HeLa cell chromosomes. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4937–4941. [PMC free article] [PubMed]
  • Avrameas S, Ternynck T. Peroxidase labelled antibody and Fab conjugates with enhanced intracellular penetration. Immunochemistry. 1971 Dec;8(12):1175–1179. [PubMed]
  • Franke WW. Structure, biochemistry, and functions of the nuclear envelope. Int Rev Cytol. 1974;Suppl 4:71–236. [PubMed]
  • Ghadially FN, Bhatnagar R, Fuller JA. Waxing and waning of nuclear fibrous lamina. Arch Pathol. 1972 Oct;94(4):303–307. [PubMed]
  • Kay RR, Fraser D, Johnston IR. A method for the rapid isolation of nuclear membranes from rat liver. Characterisation of the membrane preparation and its associated DNA polymerase. Eur J Biochem. 1972 Oct 17;30(1):145–154. [PubMed]
  • Kay RR, Johnston IR. The nuclear envelope: current problems of structure and of function. Subcell Biochem. 1973;2(2):127–167. [PubMed]
  • Maul GG, Maul HM, Scogna JE, Lieberman MW, Stein GS, Hsu BY, Borun TW. Time sequence of nuclear pore formation in phytohemagglutinin-stimulated lymphocytes and in HeLa cells during the cell cycle. J Cell Biol. 1972 Nov;55(2):433–447. [PMC free article] [PubMed]
  • Miller TE, Huang CY, Pogo AO. Rat liver nuclear skeleton and ribonucleoprotein complexes containing HnRNA. J Cell Biol. 1978 Mar;76(3):675–691. [PMC free article] [PubMed]
  • Murray RG, Murray AS, Pizzo A. The fine structure of mitosis in rat thymic lymphocytes. J Cell Biol. 1965 Aug;26(2):601–619. [PMC free article] [PubMed]
  • Oryschak AF, Ghadially FN, Bhatnagar R. Nuclear fibrous lamina in the chondrocytes of articular cartilage. J Anat. 1974 Dec;118(Pt 3):511–515. [PMC free article] [PubMed]
  • PAPPAS GD. The fine structure of the nuclear envelope of Amoeba proteus. J Biophys Biochem Cytol. 1956 Jul 25;2(4 Suppl):431–434. [PMC free article] [PubMed]
  • ROBBINS E, GONATAS NK. THE ULTRASTRUCTURE OF A MAMMALIAN CELL DURING THE MITOTIC CYCLE. J Cell Biol. 1964 Jun;21:429–463. [PMC free article] [PubMed]
  • Singer SJ, Nicolson GL. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. [PubMed]
  • Szollosi D, Calarco PG, Donahue RP. The nuclear envelope: its breakdown and fate in mammalian oogonia and oocytes. Anat Rec. 1972 Nov;174(3):325–339. [PubMed]
  • Wunderlich F, Herlan G. Reversibly contractile nuclear matrix. Its isolation, structure, and composition. J Cell Biol. 1977 May;73(2):271–278. [PMC free article] [PubMed]

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