Skip to main content
SpringerLink
Log in

Pontin is localized in nucleolar fibrillar centers

  • Research Article
  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

Pontin is a multifunctional protein having roles in various cellular processes including regulation of gene expression. Here, we addressed Pontin intracellular localization using two different monoclonal antibodies directed against different Pontin epitopes. For the first time, Pontin was directly visualized in nucleoli where it co-localizes with Upstream Binding Factor and RNA polymerase I. Nucleolar localization of Pontin was confirmed by its detection in nucleolar extracts and by electron microscopy, which revealed Pontin accumulation specifically in the nucleolar fibrillar centers. Pontin localization in the nucleolus was dynamic and Pontin accumulated in large nucleolar dots mainly during S-phase. Pontin concentration in the large nucleolar dots correlated with reduced transcriptional activity of nucleoli. In addition, Pontin was found to associate with RNA polymerase I and to interact in a complex with c-Myc with rDNA sequences indicating that Pontin is involved in the c-Myc-dependent regulation of rRNA synthesis.

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
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Andersen JS, Lyon CE, Fox AH, Leung AK, Lam YW, Steen H, Mann M, Lamond AI (2002) Directed proteomic analysis of the human nucleolus. Curr Biol 12:1–11

    Article  PubMed  Google Scholar 

  • Arabi A, Wu S, Ridderstrale K, Bierhoff H, Shiue C, Fatyol K, Fahlen S, Hydbring P, Soderberg O, Grummt I, Larsson LG, Wright AP (2005) c-Myc associates with ribosomal DNA and activates RNA polymerase I transcription. Nat Cell Biol 7:303–310

    Article  CAS  PubMed  Google Scholar 

  • Bauer A, Huber O, Kemler R (1998) Pontin52, an interaction partner of beta-catenin, binds to the TATA box binding protein. Proc Natl Acad Sci USA 95:14787–14792

    Article  CAS  PubMed  Google Scholar 

  • Bauer A, Chauvet S, Huber O, Usseglio F, Rothbacher U, Aragnol D, Kemler R, Pradel J (2000) Pontin52 and reptin52 function as antagonistic regulators of beta-catenin signalling activity. Embo J 19:6121–6130

    Article  CAS  PubMed  Google Scholar 

  • Bellosta P, Hulf T, Balla Diop S, Usseglio F, Pradel J, Aragnol D, Gallant P (2005) Myc interacts genetically with Tip48/Reptin and Tip49/Pontin to control growth and proliferation during Drosophila development. Proc Natl Acad Sci USA 102:11799–11804

    Article  CAS  PubMed  Google Scholar 

  • Draberova E, Draber P, Havlicek F, Viklicky V (1986) A common antigenic determinant of vimentin and desmin defined by monoclonal antibody. Folia Biol (Praha) 32:295–303

    CAS  Google Scholar 

  • Dugan KA, Wood MA, Cole MD (2002) TIP49, but not TRRAP, modulates c-Myc and E2F1 dependent apoptosis. Oncogene 21:5835–5843

    Article  CAS  PubMed  Google Scholar 

  • Elbashir SM, Harborth J, Weber K, Tuschl T (2002) Analysis of gene function in somatic mammalian cells using small interfering RNAs. Methods 26:199–213

    Article  CAS  PubMed  Google Scholar 

  • Etard C, Gradl D, Kunz M, Eilers M, Wedlich D (2005) Pontin and Reptin regulate cell proliferation in early Xenopus embryos in collaboration with c-Myc and Miz-1. Mech Dev 122:545–556

    Article  CAS  PubMed  Google Scholar 

  • Frank SR, Parisi T, Taubert S, Fernandez P, Fuchs M, Chan HM, Livingston DM, Amati B (2003) MYC recruits the TIP60 histone acetyltransferase complex to chromatin. EMBO Rep 4:575–580

    Article  CAS  PubMed  Google Scholar 

  • Gallant P (2007) Control of transcription by Pontin and Reptin. Trends Cell Biol 17:187–192

    Article  CAS  PubMed  Google Scholar 

  • Gartner W, Rossbacher J, Zierhut B, Daneva T, Base W, Weissel M, Waldhausl W, Pasternack MS, Wagner L (2003) The ATP-dependent helicase RUVBL1/TIP49a associates with tubulin during mitosis. Cell Motil Cytoskeleton 56:79–93

    Article  CAS  PubMed  Google Scholar 

  • Grandori C, Gomez-Roman N, Felton-Edkins ZA, Ngouenet C, Galloway DA, Eisenman RN, White RJ (2005) c-Myc binds to human ribosomal DNA and stimulates transcription of rRNA genes by RNA polymerase I. Nat Cell Biol 7:311–318

    Article  CAS  PubMed  Google Scholar 

  • Holzmann K, Gerner C, Korosec T, Poltl A, Grimm R, Sauermann G (1998) Identification and characterization of the ubiquitously occurring nuclear matrix protein NMP 238. Biochem Biophys Res Commun 252:39–45

    Article  CAS  PubMed  Google Scholar 

  • King TH, Decatur WA, Bertrand E, Maxwell ES, Fournier MJ (2001) A well-connected and conserved nucleoplasmic helicase is required for production of box C/D and H/ACA snoRNAs and localization of snoRNP proteins. Mol Cell Biol 21:7731–7746

    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 

  • Koberna K, Ligasova A, Malinsky J, Pliss A, Siegel AJ, Cvackova Z, Fidlerova H, Masata M, Fialova M, Raska I, Berezney R (2005) Electron microscopy of DNA replication in 3-D: evidence for similar-sized replication foci throughout S-phase. J Cell Biochem 94:126–138

    Article  CAS  PubMed  Google Scholar 

  • Leung AK, Trinkle-Mulcahy L, Lam YW, Andersen JS, Mann M, Lamond AI (2006) NOPdb: Nucleolar Proteome Database. Nucleic Acids Res 34:D218–D220

    Article  CAS  PubMed  Google Scholar 

  • Makino Y, Mimori T, Koike C, Kanemaki M, Kurokawa Y, Inoue S, Kishimoto T, Tamura T (1998) TIP49, homologous to the bacterial DNA helicase RuvB, acts as an autoantigen in human. Biochem Biophys Res Commun 245:819–823

    Article  CAS  PubMed  Google Scholar 

  • Malinsky J, Koberna K, Bednar J, Stulik J, Raska I (2002) Searching for active ribosomal genes in situ: light microscopy in light of the electron beam. J Struct Biol 140:227–231

    Article  CAS  PubMed  Google Scholar 

  • McKeegan KS, Debieux CM, Boulon S, Bertrand E, Watkins NJ (2007) A dynamic scaffold of pre-snoRNP factors facilitates human box C/D snoRNP assembly. Mol Cell Biol 27:6782–6793

    Article  CAS  PubMed  Google Scholar 

  • Meyer zum Buschenfelde D, Tauber R, Huber O (2006) TFF3-peptide increases transepithelial resistance in epithelial cells by modulating claudin-1 and -2 expression. Peptides 27:3383–3390

    Article  PubMed  Google Scholar 

  • Raska I (2003) Oldies but goldies: searching for Christmas trees within the nucleolar architecture. Trends Cell Biol 13:517–525

    Article  CAS  PubMed  Google Scholar 

  • Raska I, Dundr M, Koberna K, Melcak I, Risueno MC, Torok I (1995) Does the synthesis of ribosomal RNA take place within nucleolar fibrillar centers or dense fibrillar components? A critical appraisal. J Struct Biol 114:1–22

    Article  CAS  PubMed  Google Scholar 

  • Raska I, Shaw PJ, Cmarko D (2006a) New insights into nucleolar architecture and activity. Int Rev Cytol 255:177–235

    Article  CAS  PubMed  Google Scholar 

  • Raska I, Shaw PJ, Cmarko D (2006b) Structure and function of the nucleolus in the spotlight. Curr Opin Cell Biol 18:325–334

    Article  CAS  PubMed  Google Scholar 

  • Salzer U, Kubicek M, Prohaska R (1999) Isolation, molecular characterization, and tissue-specific expression of ECP-51 and ECP-54 (TIP49), two homologous, interacting erythroid cytosolic proteins. Biochim Biophys Acta 1446:365–370

    CAS  PubMed  Google Scholar 

  • Stanek D, Rader SD, Klingauf M, Neugebauer KM (2003) Targeting of U4/U6 small nuclear RNP assembly factor SART3/p110 to Cajal bodies. J Cell Biol 160:505–516

    Article  CAS  PubMed  Google Scholar 

  • Taubert S, Gorrini C, Frank SR, Parisi T, Fuchs M, Chan HM, Livingston DM, Amati B (2004) E2F-dependent histone acetylation and recruitment of the Tip60 acetyltransferase complex to chromatin in late G1. Mol Cell Biol 24:4546–4556

    Article  CAS  PubMed  Google Scholar 

  • Venteicher AS, Meng Z, Mason PJ, Veenstra TD, Artandi SE (2008) Identification of ATPases pontin and reptin as telomerase components essential for holoenzyme assembly. Cell 132:945–957

    Article  CAS  PubMed  Google Scholar 

  • Watkins NJ, Dickmanns A, Luhrmann R (2002) Conserved stem II of the box C/D motif is essential for nucleolar localization and is required, along with the 15.5K protein, for the hierarchical assembly of the box C/D snoRNP. Mol Cell Biol 22:8342–8352

    Article  CAS  PubMed  Google Scholar 

  • Watkins NJ, Lemm I, Ingelfinger D, Schneider C, Hossbach M, Urlaub H, Luhrmann R (2004) Assembly and maturation of the U3 snoRNP in the nucleoplasm in a large dynamic multiprotein complex. Mol Cell 16:789–798

    Article  CAS  PubMed  Google Scholar 

  • Weiske J, Huber O (2005) The histidine triad protein Hint1 interacts with Pontin and Reptin and inhibits TCF-beta-catenin-mediated transcription. J Cell Sci 118:3117–3129

    Article  CAS  PubMed  Google Scholar 

  • Weiske J, Huber O (2006) The histidine triad protein Hint1 triggers apoptosis independent of its enzymatic activity. J Biol Chem 281:27356–27366

    Article  CAS  PubMed  Google Scholar 

  • Wood MA, McMahon SB, Cole MD (2000) An ATPase/helicase complex is an essential cofactor for oncogenic transformation by c-Myc. Mol Cell 5:321–330

    Article  CAS  PubMed  Google Scholar 

  • Yang JM, Baserga SJ, Turley SJ, Pollard KM (2001) Fibrillarin and other snoRNP proteins are targets of autoantibodies in xenobiotic-induced autoimmunity. Clin Immunol 101:38–50

    Article  CAS  PubMed  Google Scholar 

  • Zaros C, Briand JF, Boulard Y, Labarre-Mariotte S, Garcia-Lopez MC, Thuriaux P, Navarro F (2007) Functional organization of the Rpb5 subunit shared by the three yeast RNA polymerases. Nucleic Acids Res 35:634–647

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We would like to thank Pavel Draber, Marvin Fritzer and Kenneth M. Pollard for providing us with antibodies, Klaus Holzmann (University of Vienna, Vienna, Austria) for Pontin-GFP constructs and Jasper Manning for comments on the manuscript. This work was supported by grants from the Grant Agency of the Czech Republic 301/05/0601 (to D.S.), 304/05/0374 (to K.K.), the Czech Ministry of Education (MSM0021620806, 1K05009 and LC535 to I.R.), the Wellcome trust grant 075834/04/Z (to I.R.) and the institutional projects AV0Z50520514, AV0Z50110509 and AV0Z50390512 awarded by the Academy of Sciences of the Czech Republic. The work of O.H. was supported by grants SFB366/C12 and HU881/5-1 of the Deutsche Forschungsgemeinschaft, LOM grants of the Charité—Universitätsmedizin Berlin and the Sonnenfeld Stiftung. Z.C. was supported by a grant from the Grant Agency of the Czech Republic 303/03/H065 and A.L. by 204/07/0133.

Author information

Authors and Affiliations

  1. Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Albertov 4, 128 00, Prague 2, Czech Republic

    Zuzana Cvačková, Karel Koberna, Anna Ligasová & Ivan Raška

  2. Department of Cell Biology, Institute of Physiology AS CR v.v.i., Albertov 4, 128 00, Prague 2, Czech Republic

    Zuzana Cvačková, Karel Koberna, Anna Ligasová & Ivan Raška

  3. Institute of Laboratory Medicine and Pathobiochemistry, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12200, Berlin, Germany

    Kai F. Albring & Otmar Huber

  4. Laboratory of Cell Biology, Institute of Experimental Medicine AS CR v.v.i., Vídeňská 1083, 142 20, Prague 4, Czech Republic

    Karel Koberna & Anna Ligasová

  5. Department of RNA Biology, Institute of Molecular Genetics AS CR v.v.i., Vídeňská 1083, 142 20, Prague 4, Czech Republic

    David Staněk

Authors
  1. Zuzana Cvačková
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kai F. Albring
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Karel Koberna
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Anna Ligasová
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Otmar Huber
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ivan Raška
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. David Staněk
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to David Staněk.

Additional information

Communicated by G. Matera

Zuzana Cvačková and Kai F. Albring contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cvačková, Z., Albring, K.F., Koberna, K. et al. Pontin is localized in nucleolar fibrillar centers. Chromosoma 117, 487–497 (2008). https://doi.org/10.1007/s00412-008-0170-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00412-008-0170-8

Keywords

Search

Navigation