Nucleic Acids Research, 1994, Vol. 22, No. 11 2094-2101
© 1994
MOLECULAR BIOLOGY |
Nonhomologous DNA end joining in Schizosaccharomyces pombe efficiently eliminates DNA double-strand-breaks from haploid sequences
Insitut for Genetics Weyertal, 121, 50931 Cologne, Germany
*To whom correspondence should be addressed at: Institut Curie, Pavilion Troullet-Rossignol, 26 rue d'Ulm, 75231 Paris, France
Received February 11, 1994. Accepted May 5, 1994.
Cells of higher eucaryotes are known to possess mechanisms of illegitimate recombination which promote the joining between nonhomologous ends of broken DNA and thus may serve as basic tools of double-strand-break (DSB) repair. Here we show that cells of the fission yeast Schizosaccharomyces pombe also contain activities of nonhomologous DNA end joining resembling the ones found in higher eucaryotes. Nonhomologous end joining activities were detected by transformation of linearized self-replicating plasmids in yeast cells employing a selection procedure which only propagates transformants carrying recircularized plasmid molecules. Linear plasmid substrates were generated by duplicate restriction cuts carrying either blunt ends or 3' or 5' protruding single strands (PSS) of 4 nt which were efficiently joined in any tested combination.Sequence analysis of joined products revealed that junctional sequences were shortened by 1 to 14 nt. Two mechanisms may account for junction formation (i) loss of terminal nucleotides from PSS tails to produce blunt ends which can be joined to abutting ends and (ii) interactions of DNA termini at patches of sequence homologies (1–4 bp) by formation of overlap intermediates which are subsequently processed. A general feature of the yeast joining system is that end joining can only be detected in the absence of sequence homology between the linear substrate and host genome. In the presence of homology, nonhomologous DNA end joining is efficiently competed by activities of homologous recombination.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  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]
|
|
|
|
|
|
G. Chua, L. Taricani, W. Stangle, and P. G. Young Insertional mutagenesis based on illegitimate recombination in Schizosaccharomyces pombe Nucleic Acids Res., June 1, 2000; 28(11): e53 - e53. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Manivasakam and R. H. Schiestl Nonhomologous End Joining during Restriction Enzyme-Mediated DNA Integration in Saccharomyces cerevisiae Mol. Cell. Biol., March 1, 1998; 18(3): 1736 - 1745. [Abstract] [Full Text]
|
|
|
|
 |
|
 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]
|
|
|
|
|
|
X.-Y. Gu, R. A.O. Bennett, and L. F. Povirk 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., August 16, 1996; 271(33): 19660 - 19663. [Abstract] [Full Text] [PDF]
|
|