This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Singleton, B. K.
Right arrow Articles by Jeggo, P. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Singleton, B. K.
Right arrow Articles by Jeggo, P. A.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, May 1999, p. 3267-3277, Vol. 19, No. 5
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The C Terminus of Ku80 Activates the DNA-Dependent Protein Kinase Catalytic Subunit

B. K. Singleton,1 M. I. Torres-Arzayus,2 S. T. Rottinghaus,3 G. E. Taccioli,2 and P. A. Jeggo1,*

MRC Cell Mutation Unit, University of Sussex, Brighton BN1 9RR,1 and Wellcome CRC Institute and Department of Zoology, Cambridge University, Cambridge CB2 1QR,3 United Kingdom, and Department of Microbiology, School of Medicine, Boston University, Boston, Massachusetts 021182

Received 17 August 1998/Returned for modification 27 November 1998/Accepted 22 January 1999

Ku is a heterodimeric protein with double-stranded DNA end-binding activity that operates in the process of nonhomologous end joining. Ku is thought to target the DNA-dependent protein kinase (DNA-PK) complex to the DNA and, when DNA bound, can interact and activate the DNA-PK catalytic subunit (DNA-PKcs). We have carried out a 3' deletion analysis of Ku80, the larger subunit of Ku, and shown that the C-terminal 178 amino acid residues are dispensable for DNA end-binding activity but are required for efficient interaction of Ku with DNA-PKcs. Cells expressing Ku80 proteins that lack the terminal 178 residues have low DNA-PK activity, are radiation sensitive, and can recombine the signal junctions but not the coding junctions during V(D)J recombination. These cells have therefore acquired the phenotype of mouse SCID cells despite expressing DNA-PKcs protein, suggesting that an interaction between DNA-PKcs and Ku, involving the C-terminal region of Ku80, is required for DNA double-strand break rejoining and coding but not signal joint formation. To gain further insight into important domains in Ku80, we report a point mutational change in Ku80 in the defective xrs-2 cell line. This residue is conserved among species and lies outside of the previously reported Ku70-Ku80 interaction domain. The mutational change nonetheless abrogates the Ku70-Ku80 interaction and DNA end-binding activity.

* Corresponding author. Mailing address: MRC Cell Mutation Unit, University of Sussex, Falmer, Brighton BN1 9RR, United Kingdom. Phone: 44 1273 678482. Fax: 44 1273 678121. E-mail: p.a.jeggo{at}sussex.ac.uk.

Molecular and Cellular Biology, May 1999, p. 3267-3277, Vol. 19, No. 5
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Weterings, E., Verkaik, N. S., Keijzers, G., Florea, B. I., Wang, S.-Y., Ortega, L. G., Uematsu, N., Chen, D. J., van Gent, D. C. (2009). The Ku80 Carboxy Terminus Stimulates Joining and Artemis-Mediated Processing of DNA Ends. Mol. Cell. Biol. 29: 1134-1142 [Abstract] [Full Text]  
  • Postow, L., Ghenoiu, C., Woo, E. M., Krutchinsky, A. N., Chait, B. T., Funabiki, H. (2008). Ku80 removal from DNA through double strand break-induced ubiquitylation. JCB 182: 467-479 [Abstract] [Full Text]  
  • Uematsu, N., Weterings, E., Yano, K.-i., Morotomi-Yano, K., Jakob, B., Taucher-Scholz, G., Mari, P.-O., van Gent, D. C., Chen, B. P.C., Chen, D. J. (2007). Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks. JCB 177: 219-229 [Abstract] [Full Text]  
  • Idogawa, M., Masutani, M., Shitashige, M., Honda, K., Tokino, T., Shinomura, Y., Imai, K., Hirohashi, S., Yamada, T. (2007). Ku70 and Poly(ADP-Ribose) Polymerase-1 Competitively Regulate {beta}-Catenin and T-Cell Factor-4-Mediated Gene Transactivation: Possible Linkage of DNA Damage Recognition and Wnt Signaling. Cancer Res. 67: 911-918 [Abstract] [Full Text]  
  • Niewolik, D., Pannicke, U., Lu, H., Ma, Y., Wang, L.-C. V., Kulesza, P., Zandi, E., Lieber, M. R., Schwarz, K. (2006). DNA-PKcs Dependence of Artemis Endonucleolytic Activity, Differences between Hairpins and 5' or 3' Overhangs. J. Biol. Chem. 281: 33900-33909 [Abstract] [Full Text]  
  • Drouet, J., Frit, P., Delteil, C., de Villartay, J.-P., Salles, B., Calsou, P. (2006). Interplay between Ku, Artemis, and the DNA-dependent Protein Kinase Catalytic Subunit at DNA Ends. J. Biol. Chem. 281: 27784-27793 [Abstract] [Full Text]  
  • Andrews, B. J., Lehman, J. A., Turchi, J. J. (2006). Kinetic Analysis of the Ku-DNA Binding Activity Reveals a Redox-dependent Alteration in Protein Structure That Stimulates Dissociation of the Ku-DNA Complex. J. Biol. Chem. 281: 13596-13603 [Abstract] [Full Text]  
  • Dmitrieva, N. I., Celeste, A., Nussenzweig, A., Burg, M. B. (2005). Ku86 preserves chromatin integrity in cells adapted to high NaCl. Proc. Natl. Acad. Sci. USA 102: 10730-10735 [Abstract] [Full Text]  
  • Kato, M., Nonaka, T., Imajoh-Ohmi, S. (2005). Cleavage at the Carboxyl-Terminus of Ku80 during Apoptosis in Human Jurkat T Cells. J Biochem 137: 685-692 [Abstract] [Full Text]  
  • Bladen, C. L., Lam, W. K., Dynan, W. S., Kozlowski, D. J. (2005). DNA damage response and Ku80 function in the vertebrate embryo. Nucleic Acids Res 33: 3002-3010 [Abstract] [Full Text]  
  • Lucero, H., Gae, D., Taccioli, G. E. (2003). Novel Localization of the DNA-PK Complex in Lipid Rafts: A PUTATIVE ROLE IN THE SIGNAL TRANSDUCTION PATHWAY OF THE IONIZING RADIATION RESPONSE. J. Biol. Chem. 278: 22136-22143 [Abstract] [Full Text]  
  • Dai, Y., Kysela, B., Hanakahi, L. A., Manolis, K., Riballo, E., Stumm, M., Harville, T. O., West, S. C., Oettinger, M. A., Jeggo, P. A. (2003). Nonhomologous end joining and V(D)J recombination require an additional factor. Proc. Natl. Acad. Sci. USA 100: 2462-2467 [Abstract] [Full Text]  
  • Mo, X., Dynan, W. S. (2002). Subnuclear Localization of Ku Protein: Functional Association with RNA Polymerase II Elongation Sites. Mol. Cell. Biol. 22: 8088-8099 [Abstract] [Full Text]  
  • Botta, E., Nardo, T., Lehmann, A. R., Egly, J.-M., Pedrini, A. M., Stefanini, M. (2002). Reduced level of the repair/transcription factor TFIIH in trichothiodystrophy. Hum Mol Genet 11: 2919-2928 [Abstract] [Full Text]  
  • Karmakar, P., Snowden, C. M., Ramsden, D. A., Bohr, V. A. (2002). Ku heterodimer binds to both ends of the Werner protein and functional interaction occurs at the Werner N-terminus. Nucleic Acids Res 30: 3583-3591 [Abstract] [Full Text]  
  • Arosio, D., Cui, S., Ortega, C., Chovanec, M., Di Marco, S., Baldini, G., Falaschi, A., Vindigni, A. (2002). Studies on the Mode of Ku Interaction with DNA. J. Biol. Chem. 277: 9741-9748 [Abstract] [Full Text]  
  • Marangoni, E., Foray, N., O'Driscoll, M., Douc-Rasy, S., Bernier, J., Bourhis, J., Jeggo, P. (2000). A Ku80 fragment with dominant negative activity imparts a radiosensitive phenotype to CHO-K1 cells. Nucleic Acids Res 28: 4778-4782 [Abstract] [Full Text]  
  • Tai, Y.-T., Teoh, G., Lin, B., Davies, F. E., Chauhan, D., Treon, S. P., Raje, N., Hideshima, T., Shima, Y., Podar, K., Anderson, K. C. (2000). Ku86 Variant Expression and Function in Multiple Myeloma Cells Is Associated with Increased Sensitivity to DNA Damage. J. Immunol. 165: 6347-6355 [Abstract] [Full Text]  
  • Little, J. B. (2000). Radiation carcinogenesis. Carcinogenesis 21: 397-404 [Abstract] [Full Text]  
  • Li, B., Comai, L. (2001). Requirements for the Nucleolytic Processing of DNA Ends by the Werner Syndrome Protein-Ku70/80 Complex. J. Biol. Chem. 276: 9896-9902 [Abstract] [Full Text]  
  • Woodard, R. L., Lee, K.-j., Huang, J., Dynan, W. S. (2001). Distinct Roles for Ku Protein in Transcriptional Reinitiation and DNA Repair. J. Biol. Chem. 276: 15423-15433 [Abstract] [Full Text]  
  • Koike, M., Shiomi, T., Koike, A. (2001). Dimerization and Nuclear Localization of Ku Proteins. J. Biol. Chem. 276: 11167-11173 [Abstract] [Full Text]  
  • Fukushima, T., Takata, M., Morrison, C., Araki, R., Fujimori, A., Abe, M., Tatsumi, K., Jasin, M., Dhar, P. K., Sonoda, E., Chiba, T., Takeda, S. (2001). Genetic Analysis of the DNA-dependent Protein Kinase Reveals an Inhibitory Role of Ku in Late S-G2 Phase DNA Double-strand Break Repair. J. Biol. Chem. 276: 44413-44418 [Abstract] [Full Text]  
  • Martensson, S., Hammarsten, O. (2002). DNA-dependent Protein Kinase Catalytic Subunit. STRUCTURAL REQUIREMENTS FOR KINASE ACTIVATION BY DNA ENDS. J. Biol. Chem. 277: 3020-3029 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»