Abstract
The cytoskeleton of senescent cells was systematically studied using senescent and young fibroblasts. In the cell senescence, skin fibroblasts extraordinarily produced vimentin in contrast to actin and tubulin, which were down-regulated. Among the focal adhesion proteins, paxillin and c-Src decreased also. Senescent cells developed a long and dense vimentin network, long and thin actin fibers, and numerous small focal contact sites, which contrasted with young cells with short and thick actin stress fibers and prominently large focal adhesions. Noticeably, senescent fibroblasts markedly produced p53 molecules and anchored them to vimentin-cytoskeleton in the cytoplasm. The vimentin-anchored p53 was detected with antibody PAb240 that specifically recognizes a conformation variant of p53. A GFP-tagged wild type p53 cDNA was expressed by transfection and shown also to be retained in the cytoplasm in senescent cells, suggesting that p53 is structurally modified to be recognized by PAb240 and anchored to vimentin filaments. We discuss the correlation of the marked alteration of cytoskeleton and senescent cells’ diminished proliferation and migration, as well as the significance of cytoskeletal anchorage of tumor suppressor p53.
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Akakura S, Yoshida M, Yoneda Y, Horinouchi S (2001) A role for Hsc70 in regulating nucleocytoplasmic transport of a temperature-sensitive p53 (p53Val-135). J Biol Chem 276:14649–14657
Allsopp RC, Vaziri H, Patterson C, Goldstein S, Younglai EV, Futsher AB, Greider CW, Harley CB (1992) Telomere length predicts replicative capacity of human fibroblasts. Proc Natl Acad Sci USA 89:10114–10118
Capetanaki Y, Smith S, Heath JP (1989) Overexpression of the vimentin gene in transgenic mice inhibit normal lens differentiation. J Cell Biol 109:1653–1664
Chen J, Jackson PK, Kirshner MW, Dutta A (1995) Separate domains of p21 involved in the inhibition of Cdk kinase and PCNA. Nature 374:386–388
Cho Y, Gorina S, Jeffrey PD, Pavletich NP (1994) Crystal structure of a p53 tumor suppressor-DNA complex: understanding tumorigenic mutations. Science 265:346–355
Clark EA, Brugge JS (1995) Integrins and signal transduction pathways: the road taken. Science 268:233–238
Dimri GP, Campisi J (1994) Altered profile of transcription factor-binding activities in senescent human fibroblasts. Exp Cell Res 212:132–140
Dimri GP, Hara E, Campisi J (1994) Regulation of two E2F-related genes in presenescent and senescent human fibroblasts. J Biol Chem 269:16180–16186
Ferbeyre G, de Stanchina E, Querido E, Baptiste N, Prives C, Lowe SW (2000) PML is induced by oncogenic ras and promotes premature senescence. Genes Dev 14:2015–2027
Giannakakou P, Sackett DL, Ward Y, Webster KR, Blagosklonny MV, Fojo T (2000) p53 is associated with cellular microtubules and is transported to the nucleus by dynein. Nat Cell Biol 2:709–717
Gilmore AP, Burrige K (1996) Regulation of vinculin binding to talin and actin by phosphatidylinositol-4,5-bisphosphate. Nature 381:531–535
Goldstein S (1990) Replicative senescence: the human fibroblasts comes of age. Science 249:1129–1133
Greenblatt MS, Bennett WP, Hollstein M, Harris CC (1994) Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. Cancer Res 54:4855–4878
Hayflick L (1965) The limited invitro lifetime of human diploid cell strains. Exp Cell Res 37:614–636
Kaplan KB, Robbins KB, Swedlow JR, Amaud M, Morgan DO, Varmus HE (1994) Association of the amino-terminal half of c-Src with focal adhesions alters their properties and is regulated by phosphorylation of tyrosine 527. EMBO J 13:4745–4756
Kazi AS, Lotfi S, Goncharova EA, Tliba O, Amrani Y, Krymskaya VP, Lazaar AL (2004) Vascular endothelial growth factor-induced secretion of fibronectin is ERK dependent. Am J Physiol Lung Cell Mol Physiol 286:L539–L545
Klotzache O, Etzrodt D, Hoenberg H, Bhon W, Deppert W (1998) Cytoplasmic retension of mutant tsp53 is dependent on an intermediate filament protein (vimentin) scaffold. Oncogene 16:3423–3434
Knippeschild U, Oren M, Deppert W (1996) Abrogation of wild-type p53 mediated growth-inhibition by nuclear exclusion. Oncogene 12:1755–1765
Kulju K, Lehman JM (1995) Increased p53 protein associated with aging in human diploid fibroblasts. Exp Cell Res 217:336–345
Lilienbaum A, Paulin D (1993) Activation of the human vimentin gene by the tax human T-cell leukemia virus 1. J Biol Chem 268:2180–2188
Murano S, Thweatt R, Shmookler Reis RJ, Jones R, Moerman EJ, Goldstein S (1991) Diverse gene sequences are overexpressed in Werner syndrome fibroblasts undergoing premature replicative senescence. Mol Cell Biol 8:3905–3914
Nikolaev AY, Li M, Puskas N, Qin J, Gu W (2003) Parc: a cytoplasmic anchor for p53. Cell 112:29–40
Nishio K, Inoue A, Qiao S, Kondo H, Mimura A (2001) Senescence and cytoskeleton: overproduction of vimentin induces senescent-like morphology in human fibroblasts. Histochem and Cell Biol 116:321–327
Noda A, Ning Y, Venable SF, Pereira-Smith OM, Smith JR (1994) Cloning of senescent cell-derived inhibitor of DNA synthesis using an expression screen. Exp Cell Res 211:90–98
Onyia JE, Halladay DL, Messina JL (1995) One of three CCArGG box/serum response elements of the beta-actin gene is an insulin-responsive element. Endocrinology 136:306–315
Perkins ND, Felzien LK, Bett JC, Leung K, Beach DH, Nabel GJ (1997) Regulation of NF-kB by cyclin-dependent kinases associated with p300 coactivator. Science 275:523–527
Ridley AJ, Hall A (1992) The small GTP-binding protein Rho regulates the assembly of focal adhesion and actin stress fibers in response to growth factors. Cell 70:389–399
Seshadri C, Campisi J (1990) Repression of c-fos transcription and an altered genetic program in senescent human fibroblasts. Science 247:205–200
Sin W-C, Chen X-Q, Leung T, Lim L (1998) RhoA-binding kinase alpha translocation is facilitated by the collapse of the vimentin intermediate filament network. Mol Cell Biol 18:6325–6339
Stein GH, Drullinger LF, Robetoye RS, Pereura-Smith OM, Smith JR (1991) Senescent cells fail to express cdc2, cycA and cycB in response to mitogen stimulation. Proc Natl Acad Sci USA 88:11012–11016
Uhrbom L, Nister M, Westermark B (1997) Induction of senescence in human malignant glioma cells by p16INK4A. Oncogene 15:505–514
Wang E (1985) Are cross-bridging structure involved in the bundle formation of intermediate filaments and the decrease in locomotion that accompany cell aging? J Cell Biol 100:1466–1473
Webley K, Bond JA, Jones CJ, Blaydes JP, Craig A, Hupp T, Wynford-Thomas D (2000) Posttranslational modifications of p53 in replicative senescence overlapping but distinct from those induced by DNA damage. Mol Cell Biol 20:2803–2808
Wu H, Lozano G (1994) NFkB activation of p53. A potential mechanism for suppressing cell growth in response to stress. J Biol Chem 269:20067–20074
Acknowledgements
K.N. expresses an appreciation to K. Naruse (Nagoya University) for the antibodies. We thank the Japanese Collection of Research Bioresources (JCRB) Cell Bank for the various human cells. This work was supported by a Grant-in-Aid for Scientific Research (to K. N.; 11670008, 13670009) of the Japan Society for the Promotion of Science.
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Nishio, K., Inoue, A. Senescence-associated alterations of cytoskeleton: extraordinary production of vimentin that anchors cytoplasmic p53 in senescent human fibroblasts. Histochem Cell Biol 123, 263–273 (2005). https://doi.org/10.1007/s00418-005-0766-5
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DOI: https://doi.org/10.1007/s00418-005-0766-5