| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print January 16, 2002
Molecular Biology, ICGEB (International Centre for Genetic Engineering and Biotechnology, Trieste, Trieste 34012
Corresponding Author: vindigni{at}icgeb.trieste.it
The Ku heterodimer plays a central role in non-homologous end-joining. The binding of recombinant Ku to DNA has been investigated by dynamic light scattering, double filter binding, fluorescence spectroscopy and band shift assays. The hydrodynamic radius of Ku in solution is 5.2 nm and does not change when a 25bp dsDNA fragment (D25) is added indicating that only one Ku molecule binds to a 25bp fragment. The dissociation constant (kd) for the binding to D25 is 3.8 ± 0.9 nM. If both ends of the substrate are closed with hairpin loops, Ku is still able to bind with little change in the kd. The kd is not affected by ATP, Mg2+ or ionic strength. However, the addition of BSA decreases the kd by two fold. DNA substrates of 50bp can bind two Ku molecules, while three molecules are bound to a 75bp substrate. Data analysis with the Hill equation yields a value of the Hill coefficient (n) close to 1 and the kd values for the binding of Ku to both ends of these substrates are the same. Thus, we demonstrate that there is no cooperative interaction among the Ku heterodimers binding longer substrates.
J. Biol. Chem, 10.1074/jbc.M111916200
Submitted on December 14, 2001
Revised on January 16, 2002
Accepted on January 16, 2002
Studies on the mode of Ku interaction with DNA
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  J. C.Y. Cheung, B. Salerno, and L. A. Hanakahi Evidence for an inositol hexakisphosphate-dependent role for Ku in mammalian nonhomologous end joining that is independent of its role in the DNA-dependent protein kinase Nucleic Acids Res., October 1, 2008; 36(17): 5713 - 5726. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. J. Andrews, J. A. Lehman, and J. J. Turchi 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., May 12, 2006; 281(19): 13596 - 13603. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Jin, J. Sepulveda, and O. R. Burrone Specific recognition of a dsDNA sequence motif by an immunoglobulin VH homodimer Protein Sci., December 1, 2004; 13(12): 3222 - 3229. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Arosio, S. Costantini, Y. Kong, and A. Vindigni Fluorescence Anisotropy Studies on the Ku-DNA Interaction: ANION AND CATION EFFECTS J. Biol. Chem., October 8, 2004; 279(41): 42826 - 42835. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Bentley, C. P. Diggle, P. Harnden, M. A. Knowles, and A. E. Kiltie DNA double strand break repair in human bladder cancer is error prone and involves microhomology-associated end-joining Nucleic Acids Res., October 5, 2004; 32(17): 5249 - 5259. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Cui, R. Klima, A. Ochem, D. Arosio, A. Falaschi, and A. Vindigni Characterization of the DNA-unwinding Activity of Human RECQ1, a Helicase Specifically Stimulated by Human Replication Protein A J. Biol. Chem., January 10, 2003; 278(3): 1424 - 1432. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Miyoshi, M. Sadaie, J. Kanoh, and F. Ishikawa Telomeric DNA Ends Are Essential for the Localization of Ku at Telomeres in Fission Yeast J. Biol. Chem., January 10, 2003; 278(3): 1924 - 1931. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Matheos, O. Novac, G. B. Price, and M. Zannis-Hadjopoulos Analysis of the DNA replication competence of the xrs-5 mutant cells defective in Ku86 J. Cell Sci., January 1, 2003; 116(1): 111 - 124. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
