Skip to main content
SpringerLink
Log in

Transcription-dependent rearrangements of actin and nuclear myosin I in the nucleolus

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

Nuclear actin and nuclear myosin I (NMI) are important players in transcription of ribosomal genes. Transcription of rDNA takes place in highly organized intranuclear compartment, the nucleolus. In this study, we characterized the localization of these two proteins within the nucleolus of HeLa cells with high structural resolution by means of electron microscopy and gold-immunolabeling. We demonstrate that both actin and NMI are localized in specific compartments within the nucleolus, and the distribution of NMI is transcription-dependent. Moreover, a pool of NMI is present in the foci containing nascent rRNA transcripts. Actin, in turn, is present both in transcriptionally active and inactive regions of the nucleolus and colocalizes with RNA polymerase I and UBF. Our data support the involvement of actin and NMI in rDNA transcription and point out to other functions of these proteins in the nucleolus, such as rRNA maturation and maintenance of nucleolar architecture.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Bell P, Mais C, McStay B, Scheer U (1997) Association of the nucleolar transcription factor UBF with the transcriptionally inactive rRNA genes of pronuclei and early Xenopus embryos. J Cell Sci 110(Pt 17):2053–2063

    CAS  PubMed  Google Scholar 

  • Boisvert FM, van Koningsbruggen S, Navascues J, Lamond AI (2007) The multifunctional nucleolus. Nat Rev Mol Cell Biol 8:574–585

    Article  CAS  PubMed  Google Scholar 

  • Cisterna B, Necchi D, Prosperi E, Biggiogera M (2006) Small ribosomal subunits associate with nuclear myosin and actin in transit to the nuclear pores. FASEB J 20:1901–1903

    Article  CAS  PubMed  Google Scholar 

  • Cisterna B, Malatesta M, Dieker J, Muller S, Prosperi E, Biggiogera M (2009) An active mechanism flanks and modulates the export of the small ribosomal subunits. Histochem Cell Biol 131:743–753

    Article  CAS  PubMed  Google Scholar 

  • 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 113:181–187

    Article  CAS  PubMed  Google Scholar 

  • Cruz JR, Moreno Diaz de la Espina S (2009) Subnuclear compartmentalization and function of actin and nuclear myosin I in plants. Chromosoma 118:193–207

    Article  CAS  PubMed  Google Scholar 

  • Fomproix N, Percipalle P (2004) An actin–myosin complex on actively transcribing genes. Exp Cell Res 294:140–148

    Article  CAS  PubMed  Google Scholar 

  • Hernandez-Verdun D (2006) Nucleolus: from structure to dynamics. Histochem Cell Biol 125:127–137

    Article  CAS  PubMed  Google Scholar 

  • Hirano T, Konoha G, Toda T, Yanagida M (1989) Essential roles of the RNA polymerase I largest subunit and DNA topoisomerases in the formation of fission yeast nucleolus. J Cell Biol 108:243–253

    Article  CAS  PubMed  Google Scholar 

  • Hozak P (1995) Catching RNA polymerase I in Flagranti: ribosomal genes are transcribed in the dense fibrillar component of the nucleolus. Exp Cell Res 216:285–289

    Article  CAS  PubMed  Google Scholar 

  • Hozak P, Cook PR, Schofer C, Mosgoller W, Wachtler F (1994a) Site of transcription of ribosomal RNA and intranucleolar structure in HeLa cells. J Cell Sci 107(Pt 2):639–648

    CAS  PubMed  Google Scholar 

  • Hozak P, Jackson DA, Cook PR (1994b) Replication factories and nuclear bodies: the ultrastructural characterization of replication sites during the cell cycle. J Cell Sci 107(Pt 8):2191–2202

    PubMed  Google Scholar 

  • Huang S (2002) Building an efficient factory: where is pre-rRNA synthesized in the nucleolus? J Cell Biol 157:739–741

    Article  CAS  PubMed  Google Scholar 

  • 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 133:225–234

    Article  CAS  PubMed  Google Scholar 

  • Koberna K, Malinsky J, Pliss A, Masata M, Vecerova J, Fialova M, Bednar J, Raska I (2002) Ribosomal genes in focus: new transcripts label the dense fibrillar components and form clusters indicative of “Christmas trees” in situ. J Cell Biol 157:743–748

    Article  CAS  PubMed  Google Scholar 

  • Kysela K, Philimonenko AA, Philimonenko VV, Janacek J, Kahle M, Hozak P (2005) Nuclear distribution of actin and myosin I depends on transcriptional activity of the cell. Histochem Cell Biol 124:347–358

    Article  CAS  PubMed  Google Scholar 

  • 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 109:111–118

    Article  CAS  PubMed  Google Scholar 

  • Mosgoeller W, Schofer C, Steiner M, Sylvester JE, Hozak P (2001) Arrangement of ribosomal genes in nucleolar domains revealed by detection of “Christmas tree” components. Histochem Cell Biol 116:495–505

    Article  CAS  PubMed  Google Scholar 

  • Nowak G, Pestic-Dragovich L, Hozak P, Philimonenko A, Simerly C, Schatten G, de Lanerolle P (1997) Evidence for the presence of myosin I in the nucleus. J Biol Chem 272:17176–17181

    Article  CAS  PubMed  Google Scholar 

  • Obrdlik A, Louvet E, Kukalev A, Naschekin D, Kiseleva E, Fahrenkrog B, Percipalle P (2010) Nuclear myosin 1 is in complex with mature rRNA transcripts and associates with the nuclear pore basket. FASEB J 24:146–157

    Google Scholar 

  • Olson MO, Hingorani K, Szebeni A (2002) Conventional and nonconventional roles of the nucleolus. Int Rev Cytol 219:199–266

    Article  CAS  PubMed  Google Scholar 

  • Pederson T, Tsai RY (2009) In search of nonribosomal nucleolar protein function and regulation. J Cell Biol 184:771–776

    Article  CAS  PubMed  Google Scholar 

  • Percipalle P, Fomproix N, Cavellan E, Voit R, Reimer G, Kruger T, Thyberg J, Scheer U, Grummt I, Farrants AK (2006) The chromatin remodelling complex WSTF-SNF2h interacts with nuclear myosin 1 and has a role in RNA polymerase I transcription. EMBO Rep 7:525–530

    CAS  PubMed  Google Scholar 

  • Perry RP, Kelley DE (1970) Inhibition of RNA synthesis by actinomycin D: characteristic dose-response of different RNA species. J Cell Physiol 76:127–139

    Article  CAS  PubMed  Google Scholar 

  • Philimonenko AA, Janacek J, Hozak P (2000) Statistical evaluation of colocalization patterns in immunogold labeling experiments. J Struct Biol 132:201–210

    Article  CAS  PubMed  Google Scholar 

  • Philimonenko VV, Zhao J, Iben S, Dingova H, Kysela K, Kahle M, Zentgraf H, Hofmann WA, de Lanerolle P, Hozak P, Grummt I (2004) Nuclear actin and myosin I are required for RNA polymerase I transcription. Nat Cell Biol 6:1165–1172

    Article  CAS  PubMed  Google Scholar 

  • Scheer U, Hock R (1999) Structure and function of the nucleolus. Curr Opin Cell Biol 11:385–390

    Article  CAS  PubMed  Google Scholar 

  • Schwarzacher HG, Mosgoeller W (2000) Ribosome biogenesis in man: current views on nucleolar structures and function. Cytogenet Cell Genet 91:243–252

    Article  CAS  PubMed  Google Scholar 

  • Schwarzacher HG, Wachtler F (1993) The nucleolus. Anat Embryol (Berl) 188:515–536

    CAS  Google Scholar 

  • Seither P, Zatsepina O, Hoffmann M, Grummt I (1997) Constitutive and strong association of PAF53 with RNA polymerase I. Chromosoma 106:216–225

    Article  CAS  PubMed  Google Scholar 

  • Sirri V, Urcuqui-Inchima S, Roussel P, Hernandez-Verdun D (2008) Nucleolus: the fascinating nuclear body. Histochem Cell Biol 129:13–31

    Article  CAS  PubMed  Google Scholar 

  • Thiry M, Lafontaine DL (2005) Birth of a nucleolus: the evolution of nucleolar compartments. Trends Cell Biol 15:194–199

    Article  CAS  PubMed  Google Scholar 

  • Trumtel S, Leger-Silvestre I, Gleizes PE, Teulieres F, Gas N (2000) Assembly and functional organization of the nucleolus: ultrastructural analysis of Saccharomyces cerevisiae mutants. Mol Biol Cell 11:2175–2189

    CAS  PubMed  Google Scholar 

  • Weil PA, Blatti SP (1976) HeLa cell deoxyribonucleic acid dependent RNA polymerases: function and properties of the class III enzymes. Biochemistry 15:1500–1509

    Article  CAS  PubMed  Google Scholar 

  • Ye J, Zhao J, Hoffmann-Rohrer U, Grummt I (2008) Nuclear myosin I acts in concert with polymeric actin to drive RNA polymerase I transcription. Genes Dev 22:322–330

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the Grant Agency of the Czech Republic (Reg. No. 204/07/1592), by the Ministry of the Youth and Sports (Reg. No. 2B06063, LC 545, and ME09101), and by the institutional grant IMG AS CR (Reg. No. AV0Z50520514). M.H. was supported by JSPS bilateral joint research project.

Author information

Authors and Affiliations

  1. Department of Biology of the Cell Nucleus, Institute of Molecular Genetics of the AS CR, v.v.i., Vídeňská 1083, 142 20, Prague 4, Czech Republic

    V. V. Philimonenko & P. Hozák

  2. Department of Biomathematics,, Institute of Physiology of the AS CR, v.v.i., Vídeňská 1083, 142 20, Prague 4, Czech Republic

    J. Janáček

  3. Laboratory of Molecular Biology, Graduate School of Agricultural Science, Tohoku University, Tsutsumidori-Amamiyamachi 1-1, Aoba-ku, Sendai, 981-8555, Japan

    M. Harata

Authors
  1. V. V. Philimonenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Janáček
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Harata
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P. Hozák
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to P. Hozák.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Philimonenko, V.V., Janáček, J., Harata, M. et al. Transcription-dependent rearrangements of actin and nuclear myosin I in the nucleolus. Histochem Cell Biol 134, 243–249 (2010). https://doi.org/10.1007/s00418-010-0732-8

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-010-0732-8

Keywords

Search

Navigation