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. 1995 Mar 2;128(6):1111–1119. doi: 10.1083/jcb.128.6.1111

Cytoskeletal control of gene expression: depolymerization of microtubules activates NF-kappa B

PMCID: PMC2120413  PMID: 7896875

Abstract

Cell shape changes exert specific effects on gene expression. It has been speculated that the cytoskeleton is responsible for converting changes in the cytoarchitecture to effects on gene transcription. However, the signal transduction pathways responsible for cytoskeletal- nuclear communication remained unknown. We now provide evidence that a variety of agents and conditions that depolymerize microtubules activate the sequence-specific transcription factor NF-kappa B and induce NF kappa B-dependent gene expression. These effects are caused by depolymerization of microtubule because they are blocked by the microtubule-stabilizing agent taxol. In nonstimulated cells, the majority of NF-kappa B resides in the cytosplasm as a complex with its inhibitor I kappa B. Upon cell stimulation, NF-kappa B translocates to the nucleus with concomitant degradation of I kappa B. We show that cold-induced depolymerization of microtubules also leads to I kappa B degradation and activation of NF-kappa B. However, the activated factor remains in the cytoplasm and translocates to the nucleus only upon warming to 37 degrees C, thus revealing two distinct steps in NF-kappa B activation. These findings establish a new role for NF-kappa B in sensing changes in the state of the cytoskeleton and converting them to changes in gene activity.

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Selected References

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  1. Allen J. N., Herzyk D. J., Wewers M. D. Colchicine has opposite effects on interleukin-1 beta and tumor necrosis factor-alpha production. Am J Physiol. 1991 Oct;261(4 Pt 1):L315–L321. doi: 10.1152/ajplung.1991.261.4.L315. [DOI] [PubMed] [Google Scholar]
  2. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  3. Baeuerle P. A. The inducible transcription activator NF-kappa B: regulation by distinct protein subunits. Biochim Biophys Acta. 1991 Apr 16;1072(1):63–80. doi: 10.1016/0304-419x(91)90007-8. [DOI] [PubMed] [Google Scholar]
  4. Beg A. A., Finco T. S., Nantermet P. V., Baldwin A. S., Jr Tumor necrosis factor and interleukin-1 lead to phosphorylation and loss of I kappa B alpha: a mechanism for NF-kappa B activation. Mol Cell Biol. 1993 Jun;13(6):3301–3310. doi: 10.1128/mcb.13.6.3301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beg A. A., Ruben S. M., Scheinman R. I., Haskill S., Rosen C. A., Baldwin A. S., Jr I kappa B interacts with the nuclear localization sequences of the subunits of NF-kappa B: a mechanism for cytoplasmic retention. Genes Dev. 1992 Oct;6(10):1899–1913. doi: 10.1101/gad.6.10.1899. [DOI] [PubMed] [Google Scholar]
  6. Ben-Ze'ev A. Animal cell shape changes and gene expression. Bioessays. 1991 May;13(5):207–212. doi: 10.1002/bies.950130502. [DOI] [PubMed] [Google Scholar]
  7. Benya P. D., Shaffer J. D. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell. 1982 Aug;30(1):215–224. doi: 10.1016/0092-8674(82)90027-7. [DOI] [PubMed] [Google Scholar]
  8. Bissell M. J., Hall H. G., Parry G. How does the extracellular matrix direct gene expression? J Theor Biol. 1982 Nov 7;99(1):31–68. doi: 10.1016/0022-5193(82)90388-5. [DOI] [PubMed] [Google Scholar]
  9. Blank V., Kourilsky P., Israël A. NF-kappa B and related proteins: Rel/dorsal homologies meet ankyrin-like repeats. Trends Biochem Sci. 1992 Apr;17(4):135–140. doi: 10.1016/0968-0004(92)90321-y. [DOI] [PubMed] [Google Scholar]
  10. Botteri F. M., Ballmer-Hofer K., Rajput B., Nagamine Y. Disruption of cytoskeletal structures results in the induction of the urokinase-type plasminogen activator gene expression. J Biol Chem. 1990 Aug 5;265(22):13327–13334. [PubMed] [Google Scholar]
  11. Brown K., Park S., Kanno T., Franzoso G., Siebenlist U. Mutual regulation of the transcriptional activator NF-kappa B and its inhibitor, I kappa B-alpha. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2532–2536. doi: 10.1073/pnas.90.6.2532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Carney D. H., Crossin K. L., Ball R., Fuller G. M., Albrecht T., Thompson W. C. Changes in the extent of microtubule assembly can regulate initiation of DNA synthesis. Ann N Y Acad Sci. 1986;466:919–932. doi: 10.1111/j.1749-6632.1986.tb38477.x. [DOI] [PubMed] [Google Scholar]
  13. Clayton D. F., Harrelson A. L., Darnell J. E., Jr Dependence of liver-specific transcription on tissue organization. Mol Cell Biol. 1985 Oct;5(10):2623–2632. doi: 10.1128/mcb.5.10.2623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Crossin K. L., Carney D. H. Evidence that microtubule depolymerization early in the cell cycle is sufficient to initiate DNA synthesis. Cell. 1981 Jan;23(1):61–71. doi: 10.1016/0092-8674(81)90270-1. [DOI] [PubMed] [Google Scholar]
  15. Crossin K. L., Carney D. H. Microtubule stabilization by taxol inhibits initiation of DNA synthesis by thrombin and by epidermal growth factor. Cell. 1981 Dec;27(2 Pt 1):341–350. doi: 10.1016/0092-8674(81)90417-7. [DOI] [PubMed] [Google Scholar]
  16. Davis J. Q., Bennett V. Brain ankyrin. A membrane-associated protein with binding sites for spectrin, tubulin, and the cytoplasmic domain of the erythrocyte anion channel. J Biol Chem. 1984 Nov 10;259(21):13550–13559. [PubMed] [Google Scholar]
  17. De Brabander M. J., Van de Veire R. M., Aerts F. E., Borgers M., Janssen P. A. The effects of methyl (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl) carbamate, (R 17934; NSC 238159), a new synthetic antitumoral drug interfering with microtubules, on mammalian cells cultured in vitro. Cancer Res. 1976 Mar;36(3):905–916. [PubMed] [Google Scholar]
  18. De Brabander M., Geuens G., Nuydens R., Willebrords R., De Mey J. Microtubule assembly in living cells after release from nocodazole block: the effects of metabolic inhibitors, taxol and PH. Cell Biol Int Rep. 1981 Sep;5(9):913–920. doi: 10.1016/0309-1651(81)90206-x. [DOI] [PubMed] [Google Scholar]
  19. De Brabander M., Geuens G., Nuydens R., Willebrords R., De Mey J. Taxol induces the assembly of free microtubules in living cells and blocks the organizing capacity of the centrosomes and kinetochores. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5608–5612. doi: 10.1073/pnas.78.9.5608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Devary Y., Rosette C., DiDonato J. A., Karin M. NF-kappa B activation by ultraviolet light not dependent on a nuclear signal. Science. 1993 Sep 10;261(5127):1442–1445. doi: 10.1126/science.8367725. [DOI] [PubMed] [Google Scholar]
  21. Ferrua B., Manie S., Doglio A., Shaw A., Sonthonnax S., Limouse M., Schaffar L. Stimulation of human interleukin 1 production and specific mRNA expression by microtubule-disrupting drugs. Cell Immunol. 1990 Dec;131(2):391–397. doi: 10.1016/0008-8749(90)90263-q. [DOI] [PubMed] [Google Scholar]
  22. Geiger B., Rosen D., Berke G. Spatial relationships of microtubule-organizing centers and the contact area of cytotoxic T lymphocytes and target cells. J Cell Biol. 1982 Oct;95(1):137–143. doi: 10.1083/jcb.95.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ghosh S., Gifford A. M., Riviere L. R., Tempst P., Nolan G. P., Baltimore D. Cloning of the p50 DNA binding subunit of NF-kappa B: homology to rel and dorsal. Cell. 1990 Sep 7;62(5):1019–1029. doi: 10.1016/0092-8674(90)90276-k. [DOI] [PubMed] [Google Scholar]
  24. Gilmore T. D., Morin P. J. The I kappa B proteins: members of a multifunctional family. Trends Genet. 1993 Dec;9(12):427–433. doi: 10.1016/0168-9525(93)90106-r. [DOI] [PubMed] [Google Scholar]
  25. Gordon S. R., Staley C. A. Role of the cytoskeleton during injury-induced cell migration in corneal endothelium. Cell Motil Cytoskeleton. 1990;16(1):47–57. doi: 10.1002/cm.970160107. [DOI] [PubMed] [Google Scholar]
  26. Grilli M., Chiu J. J., Lenardo M. J. NF-kappa B and Rel: participants in a multiform transcriptional regulatory system. Int Rev Cytol. 1993;143:1–62. doi: 10.1016/s0074-7696(08)61873-2. [DOI] [PubMed] [Google Scholar]
  27. Hansen S. K., Nerlov C., Zabel U., Verde P., Johnsen M., Baeuerle P. A., Blasi F. A novel complex between the p65 subunit of NF-kappa B and c-Rel binds to a DNA element involved in the phorbol ester induction of the human urokinase gene. EMBO J. 1992 Jan;11(1):205–213. doi: 10.1002/j.1460-2075.1992.tb05043.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Haskill S., Beg A. A., Tompkins S. M., Morris J. S., Yurochko A. D., Sampson-Johannes A., Mondal K., Ralph P., Baldwin A. S., Jr Characterization of an immediate-early gene induced in adherent monocytes that encodes I kappa B-like activity. Cell. 1991 Jun 28;65(7):1281–1289. doi: 10.1016/0092-8674(91)90022-q. [DOI] [PubMed] [Google Scholar]
  29. Hatada E. N., Naumann M., Scheidereit C. Common structural constituents confer I kappa B activity to NF-kappa B p105 and I kappa B/MAD-3. EMBO J. 1993 Jul;12(7):2781–2788. doi: 10.1002/j.1460-2075.1993.tb05939.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Henkel T., Machleidt T., Alkalay I., Krönke M., Ben-Neriah Y., Baeuerle P. A. Rapid proteolysis of I kappa B-alpha is necessary for activation of transcription factor NF-kappa B. Nature. 1993 Sep 9;365(6442):182–185. doi: 10.1038/365182a0. [DOI] [PubMed] [Google Scholar]
  31. Hiscott J., Marois J., Garoufalis J., D'Addario M., Roulston A., Kwan I., Pepin N., Lacoste J., Nguyen H., Bensi G. Characterization of a functional NF-kappa B site in the human interleukin 1 beta promoter: evidence for a positive autoregulatory loop. Mol Cell Biol. 1993 Oct;13(10):6231–6240. doi: 10.1128/mcb.13.10.6231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Hunter T., Karin M. The regulation of transcription by phosphorylation. Cell. 1992 Aug 7;70(3):375–387. doi: 10.1016/0092-8674(92)90162-6. [DOI] [PubMed] [Google Scholar]
  33. Ingber D. E. The riddle of morphogenesis: a question of solution chemistry or molecular cell engineering? Cell. 1993 Dec 31;75(7):1249–1252. doi: 10.1016/0092-8674(93)90612-t. [DOI] [PubMed] [Google Scholar]
  34. Katagiri K., Katagiri T., Kajiyama K., Yamamoto T., Yoshida T. Tyrosine-phosphorylation of tubulin during monocytic differentiation of HL-60 cells. J Immunol. 1993 Jan 15;150(2):585–593. [PubMed] [Google Scholar]
  35. Kieran M., Blank V., Logeat F., Vandekerckhove J., Lottspeich F., Le Bail O., Urban M. B., Kourilsky P., Baeuerle P. A., Israël A. The DNA binding subunit of NF-kappa B is identical to factor KBF1 and homologous to the rel oncogene product. Cell. 1990 Sep 7;62(5):1007–1018. doi: 10.1016/0092-8674(90)90275-j. [DOI] [PubMed] [Google Scholar]
  36. Lee J. S., von der Ahe D., Kiefer B., Nagamine Y. Cytoskeletal reorganization and TPA differently modify AP-1 to induce the urokinase-type plasminogen activator gene in LLC-PK1 cells. Nucleic Acids Res. 1993 Jul 25;21(15):3365–3372. doi: 10.1093/nar/21.15.3365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Lenardo M. J., Baltimore D. NF-kappa B: a pleiotropic mediator of inducible and tissue-specific gene control. Cell. 1989 Jul 28;58(2):227–229. doi: 10.1016/0092-8674(89)90833-7. [DOI] [PubMed] [Google Scholar]
  38. Lieuvin A., Labbé J. C., Dorée M., Job D. Intrinsic microtubule stability in interphase cells. J Cell Biol. 1994 Mar;124(6):985–996. doi: 10.1083/jcb.124.6.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Liou H. C., Baltimore D. Regulation of the NF-kappa B/rel transcription factor and I kappa B inhibitor system. Curr Opin Cell Biol. 1993 Jun;5(3):477–487. doi: 10.1016/0955-0674(93)90014-h. [DOI] [PubMed] [Google Scholar]
  40. Luduena R. F., Roach M. C. Tubulin sulfhydryl groups as probes and targets for antimitotic and antimicrotubule agents. Pharmacol Ther. 1991;49(1-2):133–152. doi: 10.1016/0163-7258(91)90027-j. [DOI] [PubMed] [Google Scholar]
  41. Lux S. E., John K. M., Bennett V. Analysis of cDNA for human erythrocyte ankyrin indicates a repeated structure with homology to tissue-differentiation and cell-cycle control proteins. Nature. 1990 Mar 1;344(6261):36–42. doi: 10.1038/344036a0. [DOI] [PubMed] [Google Scholar]
  42. Manié S., Schmid-Alliana A., Kubar J., Ferrua B., Rossi B. Disruption of microtubule network in human monocytes induces expression of interleukin-1 but not that of interleukin-6 nor tumor necrosis factor-alpha. Involvement of protein kinase A stimulation. J Biol Chem. 1993 Jun 25;268(18):13675–13681. [PubMed] [Google Scholar]
  43. Mercurio F., DiDonato J. A., Rosette C., Karin M. p105 and p98 precursor proteins play an active role in NF-kappa B-mediated signal transduction. Genes Dev. 1993 Apr;7(4):705–718. doi: 10.1101/gad.7.4.705. [DOI] [PubMed] [Google Scholar]
  44. Mitchison T. J. Mitosis: basic concepts. Curr Opin Cell Biol. 1989 Feb;1(1):67–74. doi: 10.1016/s0955-0674(89)80039-0. [DOI] [PubMed] [Google Scholar]
  45. Molony L., Armstrong L. Cytoskeletal reorganizations in human umbilical vein endothelial cells as a result of cytokine exposure. Exp Cell Res. 1991 Sep;196(1):40–48. doi: 10.1016/0014-4827(91)90454-3. [DOI] [PubMed] [Google Scholar]
  46. Murti K. G., Smith H. T., Fried V. A. Ubiquitin is a component of the microtubule network. Proc Natl Acad Sci U S A. 1988 May;85(9):3019–3023. doi: 10.1073/pnas.85.9.3019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Newmeyer D. D., Forbes D. J. Nuclear import can be separated into distinct steps in vitro: nuclear pore binding and translocation. Cell. 1988 Mar 11;52(5):641–653. doi: 10.1016/0092-8674(88)90402-3. [DOI] [PubMed] [Google Scholar]
  48. Nolan G. P., Ghosh S., Liou H. C., Tempst P., Baltimore D. DNA binding and I kappa B inhibition of the cloned p65 subunit of NF-kappa B, a rel-related polypeptide. Cell. 1991 Mar 8;64(5):961–969. doi: 10.1016/0092-8674(91)90320-x. [DOI] [PubMed] [Google Scholar]
  49. Ostlund R. E., Jr, Leung J. T., Hajek S. V. Regulation of microtubule assembly in cultured fibroblasts. J Cell Biol. 1980 May;85(2):386–391. doi: 10.1083/jcb.85.2.386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Radler-Pohl A., Pfeuffer I., Karin M., Serfling E. A novel T-cell trans-activator that recognizes a phorbol ester-inducible element of the interleukin-2 promoter. New Biol. 1990 Jun;2(6):566–573. [PubMed] [Google Scholar]
  51. Rice N. R., Ernst M. K. In vivo control of NF-kappa B activation by I kappa B alpha. EMBO J. 1993 Dec;12(12):4685–4695. doi: 10.1002/j.1460-2075.1993.tb06157.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Richardson W. D., Mills A. D., Dilworth S. M., Laskey R. A., Dingwall C. Nuclear protein migration involves two steps: rapid binding at the nuclear envelope followed by slower translocation through nuclear pores. Cell. 1988 Mar 11;52(5):655–664. doi: 10.1016/0092-8674(88)90403-5. [DOI] [PubMed] [Google Scholar]
  53. Ruben S. M., Dillon P. J., Schreck R., Henkel T., Chen C. H., Maher M., Baeuerle P. A., Rosen C. A. Isolation of a rel-related human cDNA that potentially encodes the 65-kD subunit of NF-kappa B. Science. 1991 Mar 22;251(5000):1490–1493. doi: 10.1126/science.2006423. [DOI] [PubMed] [Google Scholar]
  54. Schmid R. M., Perkins N. D., Duckett C. S., Andrews P. C., Nabel G. J. Cloning of an NF-kappa B subunit which stimulates HIV transcription in synergy with p65. Nature. 1991 Aug 22;352(6337):733–736. doi: 10.1038/352733a0. [DOI] [PubMed] [Google Scholar]
  55. Shinohara-Gotoh Y., Nishida E., Hoshi M., Sakai H. Activation of microtubule-associated protein kinase by microtubule disruption in quiescent rat 3Y1 cells. Exp Cell Res. 1991 Mar;193(1):161–166. doi: 10.1016/0014-4827(91)90551-5. [DOI] [PubMed] [Google Scholar]
  56. Sun S. C., Ganchi P. A., Ballard D. W., Greene W. C. NF-kappa B controls expression of inhibitor I kappa B alpha: evidence for an inducible autoregulatory pathway. Science. 1993 Mar 26;259(5103):1912–1915. doi: 10.1126/science.8096091. [DOI] [PubMed] [Google Scholar]
  57. Trausch J. S., Grenfell S. J., Handley-Gearhart P. M., Ciechanover A., Schwartz A. L. Immunofluorescent localization of the ubiquitin-activating enzyme, E1, to the nucleus and cytoskeleton. Am J Physiol. 1993 Jan;264(1 Pt 1):C93–102. doi: 10.1152/ajpcell.1993.264.1.C93. [DOI] [PubMed] [Google Scholar]
  58. Urban M. B., Schreck R., Baeuerle P. A. NF-kappa B contacts DNA by a heterodimer of the p50 and p65 subunit. EMBO J. 1991 Jul;10(7):1817–1825. doi: 10.1002/j.1460-2075.1991.tb07707.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Yahara I., Harada F., Sekita S., Yoshihira K., Natori S. Correlation between effects of 24 different cytochalasins on cellular structures and cellular events and those on actin in vitro. J Cell Biol. 1982 Jan;92(1):69–78. doi: 10.1083/jcb.92.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Zabel U., Henkel T., Silva M. S., Baeuerle P. A. Nuclear uptake control of NF-kappa B by MAD-3, an I kappa B protein present in the nucleus. EMBO J. 1993 Jan;12(1):201–211. doi: 10.1002/j.1460-2075.1993.tb05646.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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