Nucleic Acids Research, 1988, Vol. 16, No. 3 907-924
© 1988
Articles |
Joining of nonhomologous DNA double strand breaks in vitro
Institut für Genetik der Universität zu Köln Weyertal 121, D-5000 Köln 41, FRG
Received October 22, 1987. Accepted December 23, 1987.
Extracts of Xenopus laevis eggs can efficiently join ends of duplex DNA that differ in structure and sequence. This was analysed by recircularisation of linear plasmid DNA molecules with dissimilar termini, generated by successive cuts with two different restriction enzymes within the pSP65 polylinker. Use of various enzymes provided blunt ended or 4 nucleotides long 3' and 5' protruding single strand (PSS) termini which were successfully joined in vitro in any tested combination. Sequence analysis of numerous junctions from cloned reaction products of 7 terminus combinations reveal: apart from very rare base exchanges and single nucleotide insertions <10% deletions (1 to 18 nucleotides long) were detected. Blunt/PSS or 3'PSS/5'PSS terminus pairs undergo simple "blunt end" joining which preserves PSS ends by fill-in. In contrast, equally polar 3'PSS/3'PSS or 5'PSS/5'PSS terminus pairs are joined by a complex mode: PSS ends overlap by a defined number of nucleotides, set by matching basepairs. Even one basematch suffices to define the setting. This then determines the final mismatch repair and fill-in pattern. We propose that yet unknown terminal DNA-binding proteins stabilize the energetically highly unfavorable configuration of single matching basepairs and help to support defined overlap structures.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  S. Kuhfittig-Kulle, E. Feldmann, A. Odersky, A. Kuliczkowska, W. Goedecke, A. Eggert, and P. Pfeiffer The mutagenic potential of non-homologous end joining in the absence of the NHEJ core factors Ku70/80, DNA-PKcs and XRCC4-LigIV Mutagenesis, May 1, 2007; 22(3): 217 - 233. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Di Virgilio and J. Gautier Repair of double-strand breaks by nonhomologous end joining in the absence of Mre11 J. Cell Biol., December 5, 2005; 171(5): 765 - 771. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Yan, J. McCane, T. Toczylowski, and C. Chen Analysis of the Xenopus Werner syndrome protein in DNA double-strand break repair J. Cell Biol., October 24, 2005; 171(2): 217 - 227. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Wang, J. W. Lee, Y. Yu, K. Turner, Y. Zou, C. K. Jackson-Cook, and L. F. Povirk Gene rearrangements induced by the DNA double-strand cleaving agent neocarzinostatin: conservative non-homologous reciprocal exchanges in an otherwise stable genome Nucleic Acids Res., June 15, 2002; 30(12): 2639 - 2646. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Odersky, I. V. Panyutin, I. G. Panyutin, C. Schunck, E. Feldmann, W. Goedecke, R. D. Neumann, G. Obe, and P. Pfeiffer Repair of Sequence-specific 125I-induced Double-strand Breaks by Nonhomologous DNA End Joining in Mammalian Cell-free Extracts J. Biol. Chem., March 29, 2002; 277(14): 11756 - 11764. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Huang and W. S. Dynan Reconstitution of the mammalian DNA double-strand break end-joining reaction reveals a requirement for an Mre11/Rad50/NBS1-containing fraction Nucleic Acids Res., February 1, 2002; 30(3): 667 - 674. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Manivasakam, J. Aubrecht, S. Sidhom, and R. H. Schiestl Restriction enzymes increase efficiencies of illegitimate DNA integration but decrease homologous integration in mammalian cells Nucleic Acids Res., December 1, 2001; 29(23): 4826 - 4833. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Pastwa, R. D. Neumann, and T. A. Winters In vitro repair of complex unligatable oxidatively induced DNA double-strand breaks by human cell extracts Nucleic Acids Res., August 15, 2001; 29(16): e78 - e78. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Pospiech, A. K. Rytkonen, and J. E. Syvaoja The role of DNA polymerase activity in human non-homologous end joining Nucleic Acids Res., August 1, 2001; 29(15): 3277 - 3288. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. J. Welcker, J. de Montigny, S. Potier, and J.-L. Souciet Involvement of Very Short DNA Tandem Repeats and the Influence of the RAD52 Gene on the Occurrence of Deletions in Saccharomyces cerevisiae Genetics, October 1, 2000; 156(2): 549 - 557. [Abstract] [Full Text]
|
|
|
|
|
|
E. Feldmann, V. Schmiemann, W. Goedecke, S. Reichenberger, and P. Pfeiffer 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., July 1, 2000; 28(13): 2585 - 2596. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Pfeiffer, W. Goedecke, and G. Obe Mechanisms of DNA double-strand break repair and their potential to induce chromosomal aberrations Mutagenesis, July 1, 2000; 15(4): 289 - 302. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Tremblay, M. Jasin, and P. Chartrand 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., January 1, 2000; 20(1): 54 - 60. [Abstract] [Full Text]
|
|
|
|
|
|
P. Labhart Ku-Dependent Nonhomologous DNA End Joining in Xenopus Egg Extracts Mol. Cell. Biol., April 1, 1999; 19(4): 2585 - 2593. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. B. Cary, S. R. Peterson, J. Wang, D. G. Bear, E. M. Bradbury, and D. J. Chen DNA looping by Ku and the DNA-dependent protein kinase PNAS, April 29, 1997; 94(9): 4267 - 4272. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. S. King, C. F. Fairley, and W. F. Morgan DNA End Joining by the Klenow Fragment of DNA Polymerase I J. Biol. Chem., August 23, 1996; 271(34): 20450 - 20457. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K Meek Analysis of junctional diversity during B lymphocyte development Science, November 9, 1990; 250(4982): 820 - 823. [Abstract] [PDF]
|
|
|
|
|
|
K.-J. Lee, J. Huang, Y. Takeda, and W. S. Dynan DNA Ligase IV and XRCC4 Form a Stable Mixed Tetramer That Functions Synergistically with Other Repair Factors in a Cell-free End-joining System J. Biol. Chem., October 27, 2000; 275(44): 34787 - 34796. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Chen, K. V. Inamdar, P. Pfeiffer, E. Feldmann, M. F. Hannah, Y. Yu, J.-W. Lee, T. Zhou, S. P. Lees-Miller, and L. F. Povirk Accurate in Vitro End Joining of a DNA Double Strand Break with Partially Cohesive 3'-Overhangs and 3'-Phosphoglycolate Termini. EFFECT OF Ku ON REPAIR FIDELITY J. Biol. Chem., June 22, 2001; 276(26): 24323 - 24330. [Abstract] [Full Text] [PDF]
|
|