Cell and Tissue Research

, Volume 336, Issue 3, pp 521–527

Nuclear transport of protein TTC4 depends on the cell cycle

  • Ruslan I. Dmitriev
  • Irina A. Okkelman
  • Roman A. Abdulin
  • Mikhail I. Shakhparonov
  • Nikolay B. Pestov
Short Communication

Abstract

TTC4 (tetratricopeptide repeat domain protein 4) is a putative tumor suppressor involved in the transformation of melanocytes. At present, the relationships between TTC4 and DNA replication proteins are largely unknown, as are the tissue distribution and subcellular localization of TTC4. Using reverse transcription with the polymerase chain reaction, we have observed that the murine TTC4 gene is ubiquitously expressed. Analysis of the TTC4 subcellular localization has shown that, upon overexpression, TTC4 localizes to the cytoplasm. Interestingly, co-expression with a known protein interaction partner, hampin/MSL1, results in the nuclear translocation of the TTC4 protein. The subcellular localization of endogenous TTC4 depends, however, on the cell cycle: it is mostly nuclear in the G1 and S phases and is evenly distributed between the nucleus and cytoplasm in G2. The nuclear transport of TTC4 is apparently a complex process dependent on interactions with other proteins during the progression of the cell cycle. Thus, the dynamic character of the nuclear accumulation of TTC4 might be a potential link with regard to its function in tumor suppression.

Keywords

TTC4 Hampin Cell cycle Mouse TPR 

Supplementary material

441_2009_785_Fig1_ESM.gif (201 kb)
Figure S1

Heterogeneity of subcellular location of endogenous TTC4 protein in asynchronous fibroblasts. Sections 1 and 3 show a typical cell with predominantly nuclear TTC4, whereas in sections 2, 4 another cell from the same culture has TTC4 evenly distributed between its nucleus and cytoplasm. 3T3 cells were fixed in cold methanol and stained with anti-TTC4 antibodies followed by Alexa Fluor 488-conjugated anti-rabbit antibodies (green fluorescence) and counterstained with DAPI (blue fluorescence). 1,2 - green channel, 3,4 - merge. Bar, 50 μm. (GIF 200 KB)

441_2009_785_Fig1_ESM.tif (5.5 mb)
High resolution image file (TIF 8.3 MB)
441_2009_785_Fig2_ESM.gif (75 kb)
Figure S2

Effect of the cell cycle on subcellular distribution of the protein TTC4. 3T3 fibroblasts were synchronized by serum starvation and subsequently incubated with serum for specified time before fixation in cold methanol and staining with anti-TTC4 antibodies. Green fluorescence - Alexa Fluor 488-conjugated anti-rabbit antibodies (TTC4), blue fluorescence in left row - DAPI (nuclei). From top to bottom: 0, 7, 10, 14 and 22 hrs incubations with serum. Bar, 50 μm. (GIF 74.9 KB)

441_2009_785_Fig2_ESM.tif (19.9 mb)
High resolution image file (TIF 19.8 MB)
441_2009_785_Fig3_ESM.gif (72 kb)
Figure S3

Absence of effect of CRM1-nuclear export inhibitor LMB on TTC4 subcellular location. Hela cells were transfected by TTC4-encoding plasmid DNAs GFP-TTC4 or TTC4-RFP and treated by 100 nM LMB for 20 min. Live cells were imaged before and after treatment. 1 - TTC4-RFP before LMB treatment; 2 - TTC4-RFP after treatment; 3 - GFP-TTC4 before treatment; 4 - GFP-TTC4 after treatment. Bar, 50 μm. (GIF 72.2 KB)

441_2009_785_Fig3_ESM.tif (7.2 mb)
High resolution image file (TIF 12.8 MB)

References

  1. Blatch GL, Lässle M (1999) The tetratricopeptide repeat: a structural motif mediating protein-protein interactions. Bioessays 21:932–939PubMedCrossRefGoogle Scholar
  2. Borlado LR, Méndez J (2008) CDC6: from DNA replication to cell cycle checkpoints and oncogenesis. Carcinogenesis 29:237–243PubMedCrossRefGoogle Scholar
  3. Crevel G, Bates H, Huikeshoven H, Cotterill S (2001) The Drosophila Dpit47 protein is a nuclear Hsp90 co-chaperone that interacts with DNA polymerase alpha. J Cell Sci 114:2015–2025PubMedGoogle Scholar
  4. Crevel G, Bennett D, Cotterill S (2008) The human TPR protein TTC4 is a putative Hsp90 co-chaperone which interacts with CDC6 and shows alterations in transformed cells. PLoS ONE 3:e0001737PubMedCrossRefGoogle Scholar
  5. Delmolino LM, Saha P, Dutta A (2001) Multiple mechanisms regulate subcellular localization of human CDC6. J Biol Chem 276:26947–26954PubMedCrossRefGoogle Scholar
  6. Dmitriev RI, Pestov NB, Korneenko TV, Gerasimova AV, Zhao H, Modianov NN, Kostina MB, Shakhparonov MI (2005) Tissue specificity of alternative splicing products of mouse mRNA encoding new protein hampin homologous to the Drosophila MSL-1 protein. Bioorg Khim 31:363–371PubMedGoogle Scholar
  7. Dmitriev RI, Pestov NB, Korneenko TV, Shakhparonov MI (2006) Intracellular location of hampin isoforms. Dokl Biochem Biophys 408:130–132PubMedCrossRefGoogle Scholar
  8. Dmitriev RI, Korneenko TV, Bessonov AA, Shakhparonov MI, Modyanov NN, Pestov NB (2007) Characterization of hampin/MSL1 as a node in the nuclear interactome. Biochem Biophys Res Commun 355:1051–1057PubMedCrossRefGoogle Scholar
  9. Easwaran HP, Leonhardt H, Cardoso MC (2005) Cell cycle markers for live cell analyses. Cell Cycle 4:453–455PubMedGoogle Scholar
  10. Fabbro M, Henderson BR (2003) Regulation of tumor suppressors by nuclear-cytoplasmic shuttling. Exp Cell Res 282:59–69PubMedCrossRefGoogle Scholar
  11. Irwin N, Walker G, Hayward N (2002) Lack of TTC4 mutations in melanoma. J Invest Dermatol 119:186–187PubMedCrossRefGoogle Scholar
  12. Miyazawa H, Izumi M, Tada S, Takada R, Masutani M, Ui M, Hanaoka F (1993) Molecular cloning of the cDNAs for the four subunits of mouse DNA polymerase alpha-primase complex and their gene expression during cell proliferation and the cell cycle. J Biol Chem 268:8111–8122PubMedGoogle Scholar
  13. Mizuno T, Ito N, Yokoi M, Kobayashi A, Tamai K, Miyazawa H, Hanaoka F (1998) The second-largest subunit of the mouse DNA polymerase alpha-primase complex facilitates both production and nuclear translocation of the catalytic subunit of DNA polymerase alpha. Mol Cell Biol 18:3552–3562PubMedGoogle Scholar
  14. Orlando DA, Lin CY, Bernard A, Wang JY, Socolar JE, Iversen ES, Hartemink AJ, Haase SB (2008) Global control of cell-cycle transcription by coupled CDK and network oscillators. Nature 453:944–947PubMedCrossRefGoogle Scholar
  15. Pemberton LF, Paschal BM (2005) Mechanisms of receptor-mediated nuclear import and nuclear export. Traffic 6:187–198PubMedCrossRefGoogle Scholar
  16. Pestov NB, Korneenko TV, Adams G, Tillekeratne M, Shakhparonov MI, Modyanov NN (2002) Nongastric H-K-ATPase in rodent prostate: lobe-specific expression and apical localization. Am J Physiol Cell Physiol 282:C907–C916PubMedGoogle Scholar
  17. Pfister S, Rea S, Taipale M, Mendrzyk F, Straub B, Ittrich C, Thuerigen O, Sinn HP, Akhtar A, Lichter P (2008) The histone acetyltransferase hMOF is frequently downregulated in primary breast carcinoma and medulloblastoma and constitutes a biomarker for clinical outcome in medulloblastoma. Int J Cancer 122:1207–1213PubMedCrossRefGoogle Scholar
  18. Poetsch M, Dittberner T, Cowell JK, Woenckhaus C (2000) TTC4, a novel candidate tumor suppressor gene at 1p31 is often mutated in malignant melanoma of the skin. Oncogene 19:5817–5820PubMedCrossRefGoogle Scholar
  19. Rice JC, Nishioka K, Sarma K, Steward R, Reinberg D, Allis CD (2002) Mitotic-specific methylation of histone H4 Lys 20 follows increased PR-Set7 expression and its localization to mitotic chromosomes. Genes Dev 16:2225–2230PubMedCrossRefGoogle Scholar
  20. Smith DF (2004) Tetratricopeptide repeat cochaperones in steroid receptor complexes. Cell Stress Chaperones 9:109–121PubMedCrossRefGoogle Scholar
  21. Smith ER, Cayrou C, Huang R, Lane WS, Côté J, Lucchesi JC (2005) A human protein complex homologous to the Drosophila MSL complex is responsible for the majority of histone H4 acetylation at lysine 16. Mol Cell Biol 25:9175–9188PubMedCrossRefGoogle Scholar
  22. Su G, Roberts T, Cowell JK (1999) TTC4, a novel human gene containing the tetratricopeptide repeat and mapping to the region of chromosome 1p31 that is frequently deleted in sporadic breast cancer. Genomics 55:157–163PubMedCrossRefGoogle Scholar
  23. Su G, Casey G, Cowell JK (2000) Genomic structure of the human tetratricopeptide repeat-containing gene, TTC4, from chromosome region 1p31 and mutation analysis in breast cancers. Int J Mol Med 5:197–200PubMedGoogle Scholar
  24. Vriz S, Lemaitre JM, Leibovici M, Thierry N, Méchali M (1992) Comparative analysis of the intracellular localization of c-Myc, c-Fos, and replicative proteins during cell cycle progression. Mol Cell Biol 12:3548–3555PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Ruslan I. Dmitriev
    • 1
  • Irina A. Okkelman
    • 1
  • Roman A. Abdulin
    • 1
  • Mikhail I. Shakhparonov
    • 1
  • Nikolay B. Pestov
    • 1
  1. 1.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of SciencesMoscowRussia

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