Summary
The telomeric poly(ADP-ribose) polymerase (PARP), tankyrase 1, modulates the impact of telomerase inhibition on human cancer cells. Thus, overexpression of tankyrase 1 in telomerase-positive cancer cells confers resistance to telomerase inhibitors, such as MST-312, whereas pharmacological inhibition of tankyrase 1 enhances telomere shortening by MST-312. These facts indicate that tankyrase 1 could be a target for telomere-directed molecular cancer therapy. Here, the authors describe a convenient method to monitor the telomeric function of tankyrase 1. This protocol takes much less time than the telomere Southern blot analysis and can be utilized as a rapid screening system for tankyrase 1 inhibitors that are effective in intact cells. For direct monitoring of tankyrase 1 PARP activity, a protocol for the in vitro enzyme assay is also described.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Referencs
Smogorzewska, A. and de Lange, T. (2004) Regulation of telomerase by telomeric proteins. Annu Rev Biochem 73, 177–208.
Seimiya, H., Muramatsu, Y., Ohishi, T. and Tsuruo, T. (2005) Tankyrase 1 as a target for telomere-directed molecular cancer therapeutics. Cancer Cell 7, 25–37.
Smith, S., Giriat, I., Schmitt, A. and de Lange, T. (1998) Tankyrase, a poly(ADP-ribose) polymerase at human telomeres. Science 282, 1484–1487.
Smith, S. and de Lange, T. (2000) Tankyrase promotes telomere elongation in human cells. Curr Biol 10, 299–1302.
Chang, W., Dynek, J. N. and Smith, S. (2003) TRF1 is degraded by ubiquitin-mediated proteolysis after release from telomeres. Genes Dev 17, 1328–1333.
Loayza, D. and de Lange, T. (2003) POT1 as a terminal transducer of TRF1 telomere length control. Nature 423, 1013–1018.
Seimiya, H., Muramatsu, Y., Smith, S. and Tsuruo, T. (2004) Functional subdomain in the ankyrin domain of tankyrase 1 required for poly(ADP-ribosyl)ation of TRF1 and telomere elongation. Mol Cell Biol 24, 1944–1955.
van Steensel, B. and de Lange, T. (1997) Control of telomere length by the human telomeric protein TRF1. Nature 385, 740–743.
Seimiya, H. and Smith, S. (2002) The telomeric poly(ADP-ribose) polymerase, tankyrase 1, contains multiple binding sites for telomeric repeat binding factor 1 (TRF1) and a novel acceptor, 182-kDa tankyrase-binding protein (TAB182). J Biol Chem 277, 14116–14126.
Cook, B. D., Dynek, J. N., Chang, W., Shostak, D. and Smith, S. (2002) A role for the related poly(ADP-ribose) polymerases tankyrase 1 and 2 at human telomeres. Mol Cell Biol 22, 332–342.
Smith, S. and de Lange, T. (1999) Cell cycle dependent localization of the telomeric PARP, tankyrase, to nuclear pore complexes and centrosomes. J Cell Sci 112, 3649–3656.
Chi, N.-W. and Lodish, H. F. (2000) Tankyrase is a Golgi-associated mitogen-activated protein kinase substrate that interacts with IRAP in GLUT4 vesicles. J Biol Chem 275, 38437–38444.
Lyons, R. J., Deane, R., Lynch, D. K., Ye, Z.-S. J., Sanderson, G. M., Eyre, H. J., Sutherland, G. R. and Daly, R. J. (2001) Identification of a novel tankyrase through its interaction with the adaptor protein Grb14. J Biol Chem 276, 17172–17180.
Ye, J. Z.-S. and de Lange, T. (2004) TIN2 is a tankyrase 1 PARP modulator in the TRF1 telomere length control complex. Nat Genet 36, 618–623.
van Steensel, B., Smogorzewska, A. and de Lange, T. (1998) TRF2 protects human telomeres from end-to-end fusions. Cell 92, 401–413.
De Rycker, M., Venkatesan, R. N., Wei, C. and Price, C. M. (2003) Vertebrate tankyrase domain structure and sterile-alpha motif (SAM) mediated multimerization. Biochem J 372, 87–96.
De Rycker, M. and Price, C. M. (2004) Tankyrase polymerization is controlled by its sterile alpha motif and poly(ADP-ribose) polymerase domains. Mol Cell Biol 24, 9802–9812.
Acknowledgments
We thank Yukiko Muramatsu and Michiko Sakamoto for technical assistance and Susan Smith for scientific advice. This work was funded in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan, and grants from the Kato Memorial Bioscience Foundation and the Vehicle Racing Commemorative Foundation, Japan.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Springer
About this protocol
Cite this protocol
Ohishi, T., Tsuruo, T., Seimiya, H. (2007). Evaluation of Tankyrase Inhibition in Whole Cells. In: Andrews, L.G., Tollefsbol, T.O. (eds) Telomerase Inhibition. Methods in Molecular Biology™, vol 405. Humana Press. https://doi.org/10.1007/978-1-60327-070-0_11
Download citation
DOI: https://doi.org/10.1007/978-1-60327-070-0_11
Publisher Name: Humana Press
Print ISBN: 978-1-58829-683-2
Online ISBN: 978-1-60327-070-0
eBook Packages: Springer Protocols