This Article
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 Derbyshire, M K
Right arrow Articles by Fishel, R
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Derbyshire, M K
Right arrow Articles by Fishel, R

 Previous Article  |  Next Article 

Mol Cell Biol. 1994 January; 14(1): 156-169

Nonhomologous recombination in human cells.

M K Derbyshire, L H Epstein, C S Young, P L Munz and R Fishel

Department of Microbiology and Molecular Genetics, University of Vermont School of Medicine, Burlington 05405.

ABSTRACT

Nonhomologous recombination (NHR) is a major pathway for the repair of chromosomal double-strand breaks in the DNA of somatic cells. In this study, a comparison was made between the nonhomologous end joining of transfected adenovirus DNA fragments in vivo and the ability of purified human proteins to catalyze nonhomologous end joining in vitro. Adenovirus DNA fragments were shown to be efficiently joined in human cells regardless of the structure of the ends. Sequence analysis of these junctions revealed that the two participating ends frequently lost nucleotides from the 3' strands at the site of the joint. To examine the biochemical basis of the end joining, nuclear extracts were prepared from a wide variety of mammalian cell lines and tested for their ability to join test plasmid substrates. Efficient ligation of the linear substrate DNA was observed, the in vitro products being similar to the in vivo products with respect to the loss of 3' nucleotides at the junction. Substantial purification of the end-joining activity was carried out with the human immature T-cell-line HPB-ALL. The protein preparation was found to join all types of linear DNA substrates containing heterologous ends with closely equivalent efficiencies. The in vitro system for end joining does not appear to contain any of the three known DNA ligases, on the basis of a number of criteria, and has been termed the NHR ligase. The enriched activity resides in a high-molecular-weight recombination complex that appears to include and require the human homologous pairing protein HPP-1 as well as the NHR ligase. Characterization of the product molecules of the NHR ligase reaction suggests that they are linear oligomers of the monomer substrate joined nonrandomly head-to-head and/or tail-to-tail. The joined ends of the products were found to be modified by a 3' exonuclease prior to ligation, and no circular DNA molecules were detected. These types of products are similar to those required for the breakage-fusion-bridge cycle, a major NHR pathway for chromosome double-strand break repair.

Mol Cell Biol. 1994 January; 14(1): 156-169




This article has been cited by other articles:

  • Chan, C. Y., Kiechle, M., Manivasakam, P., Schiestl, R. H. (2007). Ionizing radiation and restriction enzymes induce microhomology-mediated illegitimate recombination in Saccharomyces cerevisiae. Nucleic Acids Res 35: 5051-5059 [Abstract] [Full Text]  
  • Rafii, S., Lindblom, A., Reed, M., Meuth, M., Cox, A. (2003). A naturally occurring mutation in an ATP-binding domain of the recombination repair gene XRCC3 ablates its function without causing cancer susceptibility. Hum Mol Genet 12: 915-923 [Abstract] [Full Text]  
  • Okorokov, A. L., Warnock, L., Milner, J. (2002). Effect of wild-type, S15D and R175H p53 proteins on DNA end joining in vitro: potential mechanism of DNA double-strand break repair modulation. Carcinogenesis 23: 549-557 [Abstract] [Full Text]  
  • Manivasakam, P., Aubrecht, J., Sidhom, S., Schiestl, R. H. (2001). Restriction enzymes increase efficiencies of illegitimate DNA integration but decrease homologous integration in mammalian cells. Nucleic Acids Res 29: 4826-4833 [Abstract] [Full Text]  
  • Lin, Y., Waldman, A. S. (2001). Promiscuous patching of broken chromosomes in mammalian cells with extrachromosomal DNA. Nucleic Acids Res 29: 3975-3981 [Abstract] [Full Text]  
  • Lin, C.-T., Lin, W.-H., Lyu, Y. L., Whang-Peng, J. (2001). Inverted repeats as genetic elements for promoting DNA inverted duplication: implications in gene amplification. Nucleic Acids Res 29: 3529-3538 [Abstract] [Full Text]  
  • Lin, Y., Waldman, A. S. (2001). Capture of DNA Sequences at Double-Strand Breaks in Mammalian Chromosomes. Genetics 158: 1665-1674 [Abstract] [Full Text]  
  • Rattray, A. J., McGill, C. B., Shafer, B. K., Strathern, J. N. (2001). Fidelity of Mitotic Double-Strand-Break Repair in Saccharomyces cerevisiae: A Role for SAE2/COM1. Genetics 158: 109-122 [Abstract] [Full Text]  
  • Feldmann, E., Schmiemann, V., Goedecke, W., Reichenberger, S., Pfeiffer, P. (2000). DNA double-strand break repair in cell-free extracts from Ku80-deficient cells: implications for Ku serving as an alignment factor in non-homologous DNA end joining. Nucleic Acids Res 28: 2585-2596 [Abstract] [Full Text]  
  • Tevelev, A., Schatz, D. G. (2000). Intermolecular V(D)J Recombination. J. Biol. Chem. 275: 8341-8348 [Abstract] [Full Text]  
  • Tremblay, A., Jasin, M., Chartrand, P. (2000). A Double-Strand Break in a Chromosomal LINE Element Can Be Repaired by Gene Conversion with Various Endogenous LINE Elements in Mouse Cells. Mol. Cell. Biol. 20: 54-60 [Abstract] [Full Text]  
  • Lin, Y., Lukacsovich, T., Waldman, A. S. (1999). Multiple Pathways for Repair of DNA Double-Strand Breaks in Mammalian Chromosomes. Mol. Cell. Biol. 19: 8353-8360 [Abstract] [Full Text]  
  • Labhart, P. (1999). Ku-Dependent Nonhomologous DNA End Joining in Xenopus Egg Extracts. Mol. Cell. Biol. 19: 2585-2593 [Abstract] [Full Text]  
  • Manivasakam, P., Schiestl, R. H. (1998). Nonhomologous End Joining during Restriction Enzyme-Mediated DNA Integration in Saccharomyces cerevisiae. Mol. Cell. Biol. 18: 1736-1745 [Abstract] [Full Text]  
  • Gu, X.-Y., Bennett, R. A.O., Povirk, L. F. (1996). End-joining of Free Radical-mediated DNA Double-strand Breaks in Vitro Is Blocked by the Kinase Inhibitor Wortmannin at a Step Preceding Removal of Damaged 3' Termini. J. Biol. Chem. 271: 19660-19663 [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»