Ends-in vs. ends-out recombination in yeast.

Genetics ◽  
1993 ◽  
Vol 135 (4) ◽  
pp. 973-980 ◽  
Author(s):  
P J Hastings ◽  
C McGill ◽  
B Shafer ◽  
J N Strathern
Keyword(s):  
Plasmid Dna ◽  
Yeast Strain ◽  
Model System ◽  
Double Strand Breaks ◽  
Linear Region ◽  
Strand Breaks ◽  
Extreme Difference ◽  
Plasmid Recovery

Abstract Integration of linearized plasmids into yeast chromosomes has been used as a model system for the study of recombination initiated by double-strand breaks. The linearized plasmid DNA recombines efficiently into sequences homologous to the ends of the DNA. This efficient recombination occurs both for the configuration in which the break is in a contiguous region of homology (herein called the ends-in configuration) and for "omega" insertions in which plasmid sequences interrupt a linear region of homology (herein called the ends-out configuration). The requirements for integration of these two configurations are expected to be different. We compared these two processes in a yeast strain containing an ends-in target and an ends-out target for the same cut plasmid. Recovery of ends-in events exceeds ends-out events by two- to threefold. Possible causes for the origin of this small bias are discussed. The lack of an extreme difference in frequency implies that cooperativity between the two ends does not contribute to the efficiency with which cut circular plasmids are integrated. This may also be true for the repair of chromosomal double-strand breaks.

scholarly journals Fragmentation of Plasmid DNA Produced by Gamma Radiation: A Theoretical Approach

2012 ◽  
Vol 2012 ◽  
pp. 1-6
Author(s):  
R. A. S. Silva ◽  
J. D. T. Arruda-Neto ◽  
L. Nieto
Keyword(s):  
Plasmid Dna ◽  
Fragment Size ◽  
Power Laws ◽  
Double Strand Breaks ◽  
Strand Breaks ◽  
Starting Point ◽  
Dna Strands ◽  
High Let ◽  
Fragment Distribution ◽  
Dna Strand

Breaks in DNA, resulting in fragmented parts, can be produced by ionizing radiation which, in turn, is the starting point in the search for novel physical aspects of DNA strands. Double-strand breaks in particular cause disruption of the DNA strand, splitting it into several fragments. In order to study effects produced by radiation in plasmid DNA, a new simple mechanical model for this molecule is proposed. In this model, a Morse-like potential and a high-LET component are used to describe the DNA-radiation interaction. Two power laws, used to fit results of the model, suggest that, firstly, distribution of fragment size is nonextensive and, secondly, that a transition phase is present in the DNA fragment distribution pattern.

scholarly journals Direct and Auger Electron-Induced, Single- and Double-Strand Breaks on Plasmid DNA Caused by 99mTc-Labeled Pyrene Derivatives and the Effect of Bonding Distance

PLoS ONE ◽  
2016 ◽  
Vol 11 (9) ◽  
pp. e0161973 ◽  
Author(s):  
Falco Reissig ◽  
Constantin Mamat ◽  
Joerg Steinbach ◽  
Hans-Juergen Pietzsch ◽  
Robert Freudenberg ◽  
...  
Keyword(s):  
Plasmid Dna ◽  
Auger Electron ◽  
Double Strand Breaks ◽  
Strand Breaks ◽  
Pyrene Derivatives

Spatial Distribution of Radiation-Induced Double-Strand Breaks in Plasmid DNA as Resolved by Atomic Force Microscopy

2005 ◽  
Vol 164 (6) ◽  
pp. 755-765 ◽  
Author(s):  
Dalong Pang ◽  
James E. Rodgers ◽  
Barry L. Berman ◽  
Sergey Chasovskikh ◽  
Anatoly Dritschilo
Keyword(s):  
Atomic Force Microscopy ◽  
Spatial Distribution ◽  
Plasmid Dna ◽  
Double Strand Breaks ◽  
Strand Breaks ◽  
Force Microscopy ◽  
Atomic Force ◽  
Radiation Induced

Repair of double-strand breaks in plasmid DNA in the yeast Saccharomyces cerevisiae

1988 ◽  
Vol 213 (2-3) ◽  
pp. 421-424 ◽  
Author(s):  
Joseph R. Perera ◽  
Alexander V. Glasunov ◽  
Vadim M. Glaser ◽  
Alla V. Boreiko
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasmid Dna ◽  
Double Strand Breaks ◽  
Yeast Saccharomyces Cerevisiae ◽  
Strand Breaks

Repairing DNA double-strand breaks by the prokaryotic non-homologous end-joining pathway

2009 ◽  
Vol 37 (3) ◽  
pp. 539-545 ◽  
Author(s):  
Nigel C. Brissett ◽  
Aidan J. Doherty
Keyword(s):  
Molecular Mechanisms ◽  
Model System ◽  
Double Strand Breaks ◽  
Repair Pathway ◽  
Dna Double Strand Breaks ◽  
End Joining ◽  
Dsb Repair ◽  
Strand Breaks ◽  
Non Homologous End Joining ◽  
Eukaryotic Organisms

The NHEJ (non-homologous end-joining) pathway is one of the major mechanisms for repairing DSBs (double-strand breaks) that occur in genomic DNA. In common with eukaryotic organisms, many prokaryotes possess a conserved NHEJ apparatus that is essential for the repair of DSBs arising in the stationary phase of the cell cycle. Although the bacterial NHEJ complex is much more minimal than its eukaryotic counterpart, both pathways share a number of common mechanistic features. The relative simplicity of the prokaryotic NHEJ complex makes it a tractable model system for investigating the cellular and molecular mechanisms of DSB repair. The present review describes recent advances in our understanding of prokaryotic end-joining, focusing primarily on biochemical, structural and cellular aspects of the mycobacterial NHEJ repair pathway.

scholarly journals Nonhomologous End-Joining Ligation Transfers DNA Regulatory Elements between Cointroduced Plasmids

2004 ◽  
Vol 24 (19) ◽  
pp. 8323-8331 ◽  
Author(s):  
Toshio Ishikawa ◽  
Eun Jig Lee ◽  
J. Larry Jameson
Keyword(s):  
Plasmid Dna ◽  
Mammalian Cells ◽  
Transfection Efficiency ◽  
Regulatory Elements ◽  
Double Strand Breaks ◽  
Nonhomologous End Joining ◽  
Dna Molecules ◽  
End Joining ◽  
Exogenous Dna ◽  
Strand Breaks

ABSTRACT Cointroduction of plasmids into mammalian cells is commonly used to investigate transcription factor regulation of reporter genes or to normalize transfection efficiency. We report here that cotransfected DNA molecules commonly transfer enhancer elements from one plasmid to another. Using separate Renilla or Firefly luciferase reporters, we found that an estrogen response element (ERE) originally linked to one of the reporters stimulated expression of the non-ERE-containing reporter. Similar enhancer transfer was seen with the cytomegalovirus enhancer. This enhancer transfer effect was not seen when cells were transfected separately with the reporters and the extracts were then combined before luciferase assays. The degree of enhancer transfer increased with transfected plasmid concentration and was greater when linearized rather than circular plasmid DNA was used. We hypothesized that double-strand breaks and heteroligation of cointroduced DNA molecules mediated the transfer of regulatory elements from one molecule to another. PCR of transfected plasmid DNA confirmed nonhomologous end-joining (NHEJ) ligation of DNA fragments originally present in separate plasmids. The NHEJ reaction was enhanced by UV light treatment to introduce double-strand breaks, and it was greater after liposome-mediated transfection than after calcium-phosphate-mediated transfection. NHEJ also occurred after adenoviral transfer of DNA into cells. We conclude that NHEJ mediates the transfer of regulatory DNA elements among cointroduced DNA molecules. These findings indicate the need for caution when interpreting results of transfection experiments containing more than one plasmid and suggest a mechanism whereby viruses or other exogenous DNA might recombine to activate unrelated genes.

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