Histochemistry and Cell Biology

, Volume 124, Issue 5, pp 347–358

Nuclear distribution of actin and myosin I depends on transcriptional activity of the cell

  • Katarína Kyselá
  • Anatoly A. Philimonenko
  • Vlada V. Philimonenko
  • Jiří Janáček
  • Michal Kahle
  • Pavel Hozák
Original paper


As previous studies suggested, nuclear myosin I (NMI) and actin have important roles in DNA transcription. In this study, we characterized the dynamics of these two proteins during transcriptional activation in phytohemagglutinin (PHA) stimulated human lymphocytes. The stimulation led to strong up-regulation of NMI both on the mRNA and protein level, while actin was relatively stably expressed. The intranuclear distribution of actin and NMI was evaluated using immunogold labeling. In nucleoli of resting cells, actin was localized predominantly to fibrillar centers (FCs), while NMI was located mainly to the dense fibrillar component (DFC). Upon stimulation, FCs remained the main site of actin localization, however, an accumulation of both actin and NMI in the DFC and in the granular component was observed. In the nucleoplasm of resting lymphocytes, both actin and NMI were localized mostly in condensed chromatin. Following stimulation, the majority of both proteins shifted towards the decondensed chromatin. In transcriptionally active cells, both actin and NMI colocalized with nucleoplasmic transcription sites. These results demonstrate that actin and NMI are compartmentalized in the nuclei where they can dynamically translocate depending on transcriptional activity of the cells.


Cell nucleus Human lymphocytes Actin Nuclear myosin I Transcription 





Fibrillar center


Dense fibrillar component


Granular component


Nuclear myosin I


Room temperature


  1. Bas A, Forsberg G, Hammarstrom S, Hammarstrom ML (2004) Utility of the housekeeping genes 18S rRNA, beta-actin and glyceraldehyde-3-phosphate-dehydrogenase for normalization in real-time quantitative reverse transcriptase-polymerase chain reaction analysis of gene expression in human T lymphocytes. Scand J Immunol 59:566–573CrossRefPubMedGoogle Scholar
  2. Bernhard W (1969) A new staining procedure for electron microscopical cytology. J Ultrastruct Res 27:250–265CrossRefPubMedGoogle Scholar
  3. Bettinger BT, Gilbert DM, Amberg DC (2004) Actin up in the nucleus. Nat Rev Mol Cell Biol 5:410–415CrossRefPubMedGoogle Scholar
  4. Boyum A (1968) Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl 97:77–89PubMedGoogle Scholar
  5. Busch H, Smetana K. (1970) The nucleolus. Academic, New YorkGoogle Scholar
  6. Carmo-Fonseca M (2002) The contribution of nuclear compartmentalization to gene regulation. Cell 108:513–521CrossRefPubMedGoogle Scholar
  7. Cmarko D, Verschure PJ, Rothblum LI, Hernandez-Verdun D, Amalric F, van Driel R, Fakan S (2000) Ultrastructural analysis of nucleolar transcription in cells microinjected with 5-bromo-UTP. Histochem Cell Biol 113:181–187CrossRefPubMedGoogle Scholar
  8. Cremer T, Kupper K, Dietzel S, Fakan S (2004) Higher order chromatin architecture in the cell nucleus: on the way from structure to function. Biol Cell 96:555–567CrossRefPubMedGoogle Scholar
  9. Fakan S (1994) Perichromatin fibrils are in situ forms of nascent transcripts. Trends Cell Biol 4:86–90CrossRefPubMedGoogle Scholar
  10. Fomproix N, Percipalle P (2004) An actin-myosin complex on actively transcribing genes. Exp Cell Res 294:140–148CrossRefPubMedGoogle Scholar
  11. Frankel S, Sigel EA, Craig C, Elgin SC, Mooseker MS, Artavanis-Tsakonas S (1997) An actin-related protein in Drosophila colocalizes with heterochromatin protein 1 in pericentric heterochromatin. J Cell Sci 110(17):1999–2012PubMedGoogle Scholar
  12. Hendzel MJ, Bazett JD (1995) RNA polymerase II transcription and the functional organization of the mammalian cell nucleus. Chromosoma 103:509–516PubMedCrossRefGoogle Scholar
  13. Hofmann WA, Stojiljkovic L, Fuchsova B, Vargas GM, Mavrommatis E, Philimonenko V, Kyselá K, Goodrich JA, Lessard JL, Hope TJ, Hozák P, de Lanerolle P (2004) Actin is part of pre-initiation complexes and is necessary for transcription by RNA polymerase II. Nat Cell Biol 6:1094–1101CrossRefPubMedGoogle Scholar
  14. Hozák P (1995) Catching RNA polymerase I in Flagranti: ribosomal genes are transcribed in the dense fibrillar component of the nucleolus 216:285–289Google Scholar
  15. Hozák P, Novák JT, Smetana K (1989) Three-dimensional reconstructions of nucleolus-organizing regions in PHA-stimulated human lymphocytes. Biology of the Cell 66:225–233CrossRefPubMedGoogle Scholar
  16. Hozák P, Cook PR, Schofer C, Mosgoller W, Wachtler F (1994) Site of transcription of ribosomal RNA and intranucleolar structure in HeLa cells. J Cell Sci 107:639–648PubMedGoogle Scholar
  17. Hu P, Wu S, Hernandez N (2004) A role for beta-actin in RNA polymerase III transcription. Genes Dev 18:3010–3015CrossRefPubMedGoogle Scholar
  18. Huang S (2002) Building an efficient factory: where is pre-rRNA synthesized in the nucleolus? J Cell Biol 157:739–741CrossRefPubMedGoogle Scholar
  19. Jackson DA (2003) The principles of nuclear structure. Chromosome Res 11:387–401CrossRefPubMedGoogle Scholar
  20. Jackson DA, Iborra FJ, Manders EM, Cook PR (1998) Numbers and organization of RNA polymerases, nascent transcripts, and transcription units in HeLa nuclei. Mol Biol Cell 9:1523–1536PubMedGoogle Scholar
  21. Jordan P, Mannervik M, Tora L, Carmo-Fonseca M (1996) In vivo evidence that TATA-binding protein/SL1 colocalizes with UBF and RNA polymerase I when rRNA synthesis is either active or inactive. J Cell Biol 133:225–234CrossRefPubMedGoogle Scholar
  22. Kimura T, Hashimoto I, Yamamoto A, Nishikawa M, Fujisawa JI (2000) Rev-dependent association of the intron-containing HIV-1 gag mRNA with the nuclear actin bundles and the inhibition of its nucleocytoplasmic transport by latrunculin-B. Genes Cells 5:289–307CrossRefPubMedGoogle Scholar
  23. Kukalev A, Nord Y, Palmberg C, Bergman T, Percipalle P (2005) Actin and hnRNP U cooperate for productive transcription by RNA polymerase II. Nat Struct Mol Biol 12:238–244CrossRefPubMedGoogle Scholar
  24. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMedGoogle Scholar
  25. Lin JJ (1981) Monoclonal antibodies against myofibrillar components of rat skeletal muscle decorate the intermediate filaments of cultured cells. Proc Natl Acad Sci USA 78:2335–2339PubMedCrossRefGoogle Scholar
  26. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  27. Mosgoeller W, Schofer C, Wesierska-Gadek J, Steiner M, Muller M, Wachtler F (1998) Ribosomal gene transcription is organized in foci within nucleolar components. Histochem Cell Biol 109:111–118CrossRefPubMedGoogle Scholar
  28. Mosgoeller W, Schofer C, Steiner M, Sylvester JE, Hozák P (2001) Arrangement of ribosomal genes in nucleolar domains revealed by detection of “Christmas tree" components. Histochem Cell Biol 116:495–505CrossRefPubMedGoogle Scholar
  29. Nakayasu H, Ueda K (1985) Association of rapidly-labelled RNAs with actin in nuclear matrix from mouse L5178Y cells. Exp Cell Res 160:319–330CrossRefPubMedGoogle Scholar
  30. Nowak G, Pestic-Dragovich L, Hozák P, Philimonenko A, Simerly C, Schatten G, de Lanerolle P (1997) Evidence for the presence of myosin I in the nucleus. J Biol Chem 272:17176–17181CrossRefPubMedGoogle Scholar
  31. Olave IA, Reck-Peterson SL, Crabtree GR (2002) Nuclear actin and actin-related proteins in chromatin remodeling. Annu Rev Biochem 71:755–781CrossRefPubMedGoogle Scholar
  32. Olson MO, Dundr M (2005) The moving parts of the nucleolus. Histochem Cell Biol 123:203–216CrossRefPubMedGoogle Scholar
  33. Olson MO, Dundr M, Szebeni A (2000) The nucleolus: an old factory with unexpected capabilities. Trends Cell Biol 10:189–196CrossRefPubMedGoogle Scholar
  34. Pederson T, Aebi U (2002) Actin in the nucleus: what form and what for? J Struct Biol 140:3–9CrossRefPubMedGoogle Scholar
  35. Percipalle P, Zhao J, Pope B, Weeds A, Lindberg U, Daneholt B (2001) Actin bound to the heterogeneous nuclear ribonucleoprotein hrp36 is associated with Balbiani ring mRNA from the gene to polysomes. J Cell Biol 153:229–236CrossRefPubMedGoogle Scholar
  36. Percipalle P, Jonsson A, Nashchekin D, Karlsson C, Bergman T, Guialis A, Daneholt B (2002) Nuclear actin is associated with a specific subset of hnRNP A/B-type proteins. Nucleic Acids Res 30:1725–1734CrossRefPubMedGoogle Scholar
  37. Pestic-Dragovich L, Stojiljkovic L, Philimonenko AA, Nowak G, Ke Y, Settlage RE, Shabanowitz J, Hunt DF, Hozák P, de Lanerolle P (2000) A myosin I isoform in the nucleus. Science 290:337–341PubMedCrossRefGoogle Scholar
  38. Philimonenko AA, Janáček J, Hozák P (2000) Statistical evaluation of colocalization patterns in immunogold labeling experiments. J Struct Biol 132:201–210PubMedCrossRefGoogle Scholar
  39. Philimonenko VV, Zhao J, Iben S, Dingová H, Kyselá K, Kahle M, Zentgraf H, Hofmann WA, de Lanerolle P, Hozák P, Grummt I (2004) Nuclear actin and myosin I are required for RNA polymerase I transcription. Nat Cell Biol 6:1165–1172PubMedCrossRefGoogle Scholar
  40. Puvion E, Puvion-Dutilleul F (1996) Ultrastructure of the nucleus in relation to transcription and splicing: roles of perichromatin fibrils and interchromatin granules. Exp Cell Res 229:217–225PubMedCrossRefGoogle Scholar
  41. Rando OJ, Zhao K, Crabtree GR (2000) Searching for a function for nuclear actin. Trends Cell Biol 10:92–97PubMedCrossRefGoogle Scholar
  42. Roussel P, Andre C, Comai L, Hernandez-Verdun D (1996) The rDNA transcription machinery is assembled during mitosis in active NORs and absent in inactive NORs. J Cell Biol 133:235–246PubMedCrossRefGoogle Scholar
  43. Rungger D, Rungger-Brandle E, Chaponnier C, Gabbiani G (1979) Intranuclear injection of anti-actin antibodies into Xenopus oocytes blocks chromosome condensation. Nature 282:320–321PubMedCrossRefGoogle Scholar
  44. Scheer U, Hock R (1999) Structure and function of the nucleolus. Curr Opin Cell Biol 11:385–390PubMedCrossRefGoogle Scholar
  45. Schwarzacher HG, Mosgoeller W (2000) Ribosome biogenesis in man: current views on nucleolar structures and function. Cytogenet Cell Genet 91:243–252PubMedCrossRefGoogle Scholar
  46. Setterfield G, Hall R, Bladon T, Little J, Kaplan JG (1983) Changes in structure and composition of lymphocyte nuclei during mitogenic stimulation. J Ultrastruct Res 82:264–282PubMedCrossRefGoogle Scholar
  47. Sirri V, Roussel P, Hernandez-Verdun D (1999) The mitotically phosphorylated form of the transcription termination factor TTF-1 is associated with the repressed rDNA transcription machinery. J Cell Sci 112(19):3259–3268PubMedGoogle Scholar
  48. Smetana K, Potměšil M (1968) Ring shaped nucleoli in liver cells of rats after treatment with actinomycin D. Z Zellforsch Mikrosk Anat 92(1):62–69PubMedCrossRefGoogle Scholar
  49. Spector DL (2003) The dynamics of chromosome organization and gene regulation. Annu Rev Biochem 72:573–608PubMedCrossRefGoogle Scholar
  50. Steen HB, Lindmo T (1979) Initiation of the blastogenic response of lymphocytes by hyperoptimal concentrations of concanavalin A. Eur J Immunol 9:434–439PubMedCrossRefGoogle Scholar
  51. Suzuki T, Higgins PJ, Crawford DR (2000) Control selection for RNA quantitation. Biotechniques 29:332–337PubMedGoogle Scholar
  52. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354PubMedCrossRefGoogle Scholar
  53. Visintin R, Amon A (2000) The nucleolus: the magician’s hat for cell cycle tricks. Curr Opin Cell Biol 12:372–377PubMedCrossRefGoogle Scholar
  54. Wada A, Fukuda M, Mishima M, Nishida E (1998) Nuclear export of actin: a novel mechanism regulating the subcellular localization of a major cytoskeletal protein. Embo J 17:1635–1641PubMedCrossRefGoogle Scholar
  55. Zatsepina OV, Hozák P, Babadjanyan D, Chentsov Y (1988) Quantitative ultrastructural study of nucleolus-organizing regions at some stages of the cell cycle (G0 period, G2 period, mitosis). Biol Cell 62:211–218PubMedCrossRefGoogle Scholar
  56. Zhao K, Wang W, Rando OJ, Xue Y, Swiderek K, Kuo A, Crabtree GR (1998) Rapid and phosphoinositol-dependent binding of the SWI/SNF-like BAF complex to chromatin after T lymphocyte receptor signaling. Cell 95:625–636PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Katarína Kyselá
    • 1
  • Anatoly A. Philimonenko
    • 1
  • Vlada V. Philimonenko
    • 1
  • Jiří Janáček
    • 2
  • Michal Kahle
    • 1
  • Pavel Hozák
    • 1
  1. 1.Department of Cell Ultrastructure and Molecular Biology, Institute of Experimental MedicineAcademy of Sciences of the Czech RepublicPrague 4Czech Republic
  2. 2.Department of Biomathematics, Institute of PhysiologyAcademy of Sciences of the Czech RepublicPrague 4Czech Republic

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