Cytoplasmic Intermediate Filaments Are Stably Associated with Nuclear Matrices and Potentially Modulate Their DNA-Binding Function

Tolstonog, Genrich V.; Sabasch, Michael; Traub, Peter
March 2002
DNA & Cell Biology;Mar2002, Vol. 21 Issue 3, p213
Academic Journal
The tight association of cytoplasmic intermediate filaments (cIFs) with the nucleus and the isolation of crosslinkage products of vimentin with genomic DNA fragments, including nuclear matrix attachment regions (MARs) from proliferating fibroblasts, point to a participation of cIFs in nuclear activities. To test the possibility that cIFs are complementary nuclear matrix elements, the nuclei of a series of cultured cells were subjected to the Li-diiodosalicylate (LIS) extraction protocol developed for the preparation of nuclear matrices and analyzed by immunofluorescence microscopy and immunoblotting with antibodies directed against lamin B and cIF proteins. When nuclei released from hypotonically swollen L929 suspension cells in the presence of digitonin or Triton X-100 were exposed to such strong shearing forces that a considerable number were totally disrupted, a thin, discontinuous layer of vimentin IFs remained tenaciously adhering to still intact nuclei, in apparent coalignment with the nuclear lamina. Even in broken nuclei, the distribution of vimentin followed that of lamin B in areas where the lamina still appeared intact. The same retention of vimentin together with desmin and glial IFs was observed on the nuclei isolated from differentiating C2C12 myoblast and U333 glioma cells, respectively. Nuclei from epithelial cells shed their residual perinuclear IF layers as coherent cytoskeletal ghosts, except for small fractions of vimentin and cytokeratin IFs, which remained in a dot-to cap-like arrangement on the nuclear surface, in apparent codistribution with lamin B. LIS extraction did not bring about a reduction in the cIF protein contents of such nuclei upon their transformation into nuclear matrices. Moreover, in whole mount preparations of mouse embryo fibroblasts, DNA/chromatin emerging from nuclei during LIS extraction mechanically and chemically cleaned the nuclear surface and perinuclear area from loosely anchored cytoplasmic material with the production of broad, IF-free annular spaces, but left substantial fractions of the vimentin IFs in tight association with the nuclear surface. Accordingly, double-immunogold electron microscopy of fixed and permeabilized fibroblasts disclosed a close neighborhood of vimentin IFs and lamin B, with a minimal distance between the nanogold particles of ca. 30 nm. These data indicate an extremely solid interconnection of cIFs with structural elements of the nuclear matrix, and make them, together with their susceptibility to crosslinkage to MARs and other genomic DNA sequences under native conditions, complementary or even integral constituents of the karyoskeleton.


Related Articles

  • The nucleoskeleton as a genome-associated dynamic 'network of networks'. Simon, Dan N.; Wilson, Katherine L. // Nature Reviews Molecular Cell Biology;Nov2011, Vol. 12 Issue 11, p695 

    In the cytosol, actin polymers, intermediate filaments and microtubules can anchor to cell surface adhesions and interlink to form intricate networks. This cytoskeleton is anchored to the nucleus through LINC (links the nucleoskeleton and cytoskeleton) complexes that span the nuclear envelope...

  • Elicitor-Induced Cytoskeletal Rearrangement Relates to Vacuolar Dynamics and Execution of Cell Death: In Vivo Imaging of Hypersensitive Cell Death in Tobacco BY-2 Cells. Takumi Higaki; Tatsuaki Goh; Teruyuki Hayashi; Natsumaro Kutsuna; Yasuhiro Kadota; Seiichiro Hasezawa; Toshio Sano; Kazuyuki Kuchitsu // Plant & Cell Physiology;Oct2007, Vol. 48 Issue 10, p1414 

    Disintegration of the vacuolar membrane (VM) has been proposed to be a crucial event in various types of programmed cell death (PCD) in plants. However, its regulatory mechanisms are mostly unknown. To obtain new insights on the regulation of VM disintegration during hypersensitive cell death,...

  • Influence of Fimasartan (a Novel AT1 Receptor Blocker) on Catecholamine Release in the Adrenal Medulla of Spontaneously Hypertensive Rats. Hyo-Jeong Lim; Seog-Ki Lee; Dong-Yoon Lim // Korean Journal of Physiology & Pharmacology;Feb2013, Vol. 17 Issue 1, p99 

    The aim of this study was to determine whether fimasartan, a newly developed AT1 receptor blocker, can affect the CA release in the isolated perfused model of the adrenal medulla of spontaneously hypertensive rats (SHRs). Fimasartan (5∼50 μM) perfused into an adrenal vein for 90 min...

  • Coordination of microtubule and microfilament dynamics by Drosophila Rho1, Spire and Cappuccino. Rosales-Nieves, Alicia E.; Johndrow, James E.; Keller, Lani C.; Magie, Craig R.; Pinto-Santini, Delia M.; Parkhurst, Susan M. // Nature Cell Biology;Apr2006, Vol. 8 Issue 4, p367 

    The actin-nucleation factors Spire and Cappuccino (Capu) regulate the onset of ooplasmic streaming in Drosophila melanogaster. Although this streaming event is microtubule-based, actin assembly is required for its timing. It is not understood how the interaction of microtubules and...

  • Embryogenesis in Sedum acre L.: structural and immunocytochemical aspects of suspensor development. Kozieradzka-Kiszkurno, Małgorzata; Świerczyńska, Joanna; Bohdanowicz, Jerzy // Protoplasma;Oct2011, Vol. 248 Issue 4, p775 

    The changes in the formation of both the actin and the microtubular cytoskeleton during the differentiation of the embryo-suspensor in Sedum acre were studied in comparison with the development of the embryo-proper. The presence and distribution of the cytoskeletal elements were examined...

  • Insights into eisosome assembly and organization. Murphy, E.; Kim, K. // Journal of Biosciences;Jun2012, Vol. 37 Issue 2, p295 

    Eisosomes, large protein complexes that are predominantly composed of BAR-domain-containing proteins Pil1 and its homologs, are situated under the plasma membrane of ascomycetes. A successful targeting of Pil1 onto the future site of eisosome accompanies maturation of eisosome. During or after...

  • Syncoilin, an intermediate filament-like protein linked to the dystrophin associated protein complex in skeletal muscle. Moorwood, C. // Cellular & Molecular Life Sciences;Oct2008, Vol. 65 Issue 19, p2957 

    Syncoilin is a member of the intermediate filament protein family, highly expressed in skeletal and cardiac muscle. Syncoilin binds α-dystrobrevin, a component of the dystrophin associated protein complex (DAPC) located at the muscle cell membrane, and desmin, a muscle-specific intermediate...

  • Possible Involvement of Mechanosensitive Ca2+ Channels of Plasma Membrane in Mechanoperception in Chara. Kaneko, Toshiyuki; Saito, Chiyuki; Shimmen, Teruo; Kikuyama, Munehiro // Plant & Cell Physiology;Jan2005, Vol. 46 Issue 1, p130 

    When an internodal cell of Chara corallina was stimulated with a mechanical pulse of various amplitudes lasting for 0.1 s (mechanical stimulus), the cell generated a receptor potential, which was highly dependent not only on the strength of the stimulus but also on the extracellular...

  • Bacterial physiology: Remodelling the FtsZ network. Du Toit, Andrea // Nature Reviews Microbiology;Feb2014, Vol. 12 Issue 2, p77 

    The article reports on a research study which examines the formation of Z-ring and the role of bacterial cell division protein FtsA in FtsZ network. Findings of the study reveal the attachment of FtsZ filaments to the membrane during the presence of FtsA and the static condition of single...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics