Skip Navigation

This Article
Right arrow Print PDF (5278K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
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 Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (57)
Right arrowRequest Permissions
Citing Articles
Right arrowScopus Links
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Ono, M.
Right arrow Articles by Capra, J.D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ono, M.
Right arrow Articles by Capra, J.D.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1994, Vol. 22, No. 19 3918-3924
© 1994

MOLECULAR BIOLOGY

Production and characterization of recombinant human Ku antigen

Masashi Ono, Philip W. Tucker and J.Donald Capra*

Departments of Microbiology and Dermatology, University of Texas Southwestern Medical Center Dallas, TX 75235, USA

*To whom correspondence should be addressed

Received June 10, 1994. Revised August 17, 1994. Accepted August 17, 1994.

Ku is an ubiquitous nuclear heterodimeric protein consisting of p70 and p86 subunits that binds doublestranded DNA termini and associates with chromosomes in vivo. It was originally described as an autoantigen in patients with certain autoimmune diseases. The individual subunits of Ku have been difficult to isolate from human cells without denaturation and attempts to produce functional recombinant Ku have been largely unsuccessful. Here, we utilize two recombinant baculoviral vectors that carry p70 or p86 cDNA and express the Ku subunits individually as well as assemble them into the complete Ku heterodimer. In an electrophoretic mobility shift assay, recombinant Ku binds to linear double-stranded DNA but not to supercoiled, nicked circular, nor linear single-stranded DNA. Neither subunit binds DNA by itself indicating that heterodimerization is essential for function. We also describe a simple purification method for the isolation of highly purified recombinant Ku using a hexahistidine tag. The baculovirus expression system provides a stable and efficient source of not only the p70 and p86 subunits but also the functional Ku heterodimer.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
T.-J. Wu, Y.-H. Chiang, Y.-C. Lin, C.-R. Tsai, T.-Y. Yu, M.-T. Sung, Y.-H. W. Lee, and J.-J. Lin
Sequential Loading of Saccharomyces cerevisiae Ku and Cdc13p to Telomeres
J. Biol. Chem., May 8, 2009; 284(19): 12801 - 12808.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
E. Weterings, N. S. Verkaik, G. Keijzers, B. I. Florea, S.-Y. Wang, L. G. Ortega, N. Uematsu, D. J. Chen, and D. C. van Gent
The Ku80 Carboxy Terminus Stimulates Joining and Artemis-Mediated Processing of DNA Ends
Mol. Cell. Biol., March 1, 2009; 29(5): 1134 - 1142.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Drouet, P. Frit, C. Delteil, J.-P. de Villartay, B. Salles, and P. Calsou
Interplay between Ku, Artemis, and the DNA-dependent Protein Kinase Catalytic Subunit at DNA Ends
J. Biol. Chem., September 22, 2006; 281(38): 27784 - 27793.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Jovanovic and W. S. Dynan
Terminal DNA structure and ATP influence binding parameters of the DNA-dependent protein kinase at an early step prior to DNA synapsis
Nucleic Acids Res., February 18, 2006; 34(4): 1112 - 1120.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
G. L. Mayeur, W.-J. Kung, A. Martinez, C. Izumiya, D. J. Chen, and H.-J. Kung
Ku Is a Novel Transcriptional Recycling Coactivator of the Androgen Receptor in Prostate Cancer Cells
J. Biol. Chem., March 18, 2005; 280(11): 10827 - 10833.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
N. Beerens, J. H. J. Hoeijmakers, R. Kanaar, W. Vermeulen, and C. Wyman
The CSB Protein Actively Wraps DNA
J. Biol. Chem., February 11, 2005; 280(6): 4722 - 4729.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J. Byrum, S. Jordan, S. T. Safrany, and W. Rodgers
Visualization of inositol phosphate-dependent mobility of Ku: depletion of the DNA-PK cofactor InsP6 inhibits Ku mobility
Nucleic Acids Res., May 18, 2004; 32(9): 2776 - 2784.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
D. Ristic, M. Modesti, R. Kanaar, and C. Wyman
Rad52 and Ku bind to different DNA structures produced early in double-strand break repair
Nucleic Acids Res., September 15, 2003; 31(18): 5229 - 5237.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
W. Rodgers, S. J. Jordan, and J. D. Capra
Transient Association of Ku with Nuclear Substrates Characterized Using Fluorescence Photobleaching
J. Immunol., March 1, 2002; 168(5): 2348 - 2355.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
G. Achanta, H. Pelicano, L. Feng, W. Plunkett, and P. Huang
Interaction of p53 and DNA-PK in Response to Nucleoside Analogues: Potential Role As a Sensor Complex for DNA Damage
Cancer Res., December 1, 2001; 61(24): 8723 - 8729.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
O. Novac, D. Matheos, F. D. Araujo, G. B. Price, and M. Zannis-Hadjopoulos
In Vivo Association of Ku with Mammalian Origins of DNA Replication
Mol. Biol. Cell, November 1, 2001; 12(11): 3386 - 3401.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
D. Ristic, C. Wyman, C. Paulusma, and R. Kanaar
The architecture of the human Rad54-DNA complex provides evidence for protein translocation along DNA
PNAS, July 17, 2001; 98(15): 8454 - 8460.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. de Jager, M. L. G. Dronkert, M. Modesti, C. E. M. T. Beerens, R. Kanaar, and D. C. van Gent
DNA-binding and strand-annealing activities of human Mre11: implications for its roles in DNA double-strand break repair pathways
Nucleic Acids Res., March 15, 2001; 29(6): 1317 - 1325.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
T. Mizuno, K. Yamagishi, H. Miyazawa, and F. Hanaoka
Molecular Architecture of the Mouse DNA Polymerase alpha -Primase Complex
Mol. Cell. Biol., November 1, 1999; 19(11): 7886 - 7896.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
S. Mickelsen, C. Snyder, K. Trujillo, M. Bogue, D. B. Roth, and K. Meek
Modulation of Terminal Deoxynucleotidyltransferase Activity by the DNA-Dependent Protein Kinase
J. Immunol., July 15, 1999; 163(2): 834 - 843.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Yoo, A. Kimzey, and W. S. Dynan
Photocross-linking of an Oriented DNA Repair Complex. Ku BOUND AT A SINGLE DNA END
J. Biol. Chem., July 9, 1999; 274(28): 20034 - 20039.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
W. Giffin, W. Gong, C. Schild-Poulter, and R. J. G. Hache
Ku Antigen-DNA Conformation Determines the Activation of DNA-Dependent Protein Kinase and DNA Sequence-Directed Repression of Mouse Mammary Tumor Virus Transcription
Mol. Cell. Biol., June 1, 1999; 19(6): 4065 - 4078.
[Abstract] [Full Text] [PDF]

Home page
 Genes Dev.Home page
G. C.M. Smith and S. P. Jackson
The DNA-dependent protein kinase
Genes & Dev., April 15, 1999; 13(8): 916 - 934.
[Full Text]

Home page
 J. Biol. Chem.Home page
P. Calsou, P. Frit, O. Humbert, C. Muller, D. J. Chen, and B. Salles
The DNA-dependent Protein Kinase Catalytic Activity Regulates DNA End Processing by Means of Ku Entry into DNA
J. Biol. Chem., March 19, 1999; 274(12): 7848 - 7856.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. Torrance, W. Giffin, D. J. Rodda, L. Pope, and R. J. G. Hache
Sequence-specific Binding of Ku Autoantigen to Single-stranded DNA
J. Biol. Chem., August 14, 1998; 273(33): 20810 - 20819.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. Leber, T. W. Wise, R. Mizuta, and K. Meek
The XRCC4 Gene Product Is a Target for and Interacts with the DNA-dependent Protein Kinase
J. Biol. Chem., January 16, 1998; 273(3): 1794 - 1801.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Wang, X. Dong, K. Myung, E. A. Hendrickson, and W. H. Reeves
Identification of Two Domains of the p70 Ku Protein Mediating Dimerization with p80 and DNA Binding
J. Biol. Chem., January 9, 1998; 273(2): 842 - 848.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
O. Osipovich, S. K. Durum, and K. Muegge
Defining the Minimal Domain of Ku80 for Interaction with Ku70
J. Biol. Chem., October 24, 1997; 272(43): 27259 - 27265.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Huang, A. Nueda, S. Yoo, and W. S. Dynan
Heat Shock Transcription Factor 1 Binds Selectively in Vitro to Ku Protein and the Catalytic Subunit of the DNA-dependent Protein Kinase
J. Biol. Chem., October 10, 1997; 272(41): 26009 - 26016.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
W. Giffin, J. Kwast-Welfeld, D. J. Rodda, G. G. Prefontaine, M. Traykova-Andonova, Y. Zhang, N. L. Weigel, Y. A. Lefebvre, and R. J.G. Hache
Sequence-specific DNA Binding and Transcription Factor Phosphorylation by Ku Autoantigen/DNA-dependent Protein Kinase. PHOSPHORYLATION OF Ser-527 OF THE RAT GLUCOCORTICOID RECEPTOR
J. Biol. Chem., February 28, 1997; 272(9): 5647 - 5658.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. Frit, R.-Y. Li, D. Arzel, B. Salles, and P. Calsou
Ku Entry into DNA Inhibits Inward DNA Transactions in Vitro
J. Biol. Chem., November 10, 2000; 275(46): 35684 - 35691.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. L. Woodard, K.-j. Lee, J. Huang, and W. S. Dynan
Distinct Roles for Ku Protein in Transcriptional Reinitiation and DNA Repair
J. Biol. Chem., April 27, 2001; 276(18): 15423 - 15433.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.