The deinococcal DdrB protein is involved in an early step of DNA double strand break repair and in plasmid transformation through its single-strand annealing activity

ABSTRACT A number of studies of Saccharomyces cerevisiae have revealed RAD51-independent recombination events. These include spontaneous and double-strand break-induced recombination between repeated sequences, and capture of a chromosome arm by break-induced replication. Although recombination between inverted repeats is considered to be a conservative intramolecular event, the lack of requirement for RAD51 suggests that repair can also occur by a nonconservative mechanism. We propose a model forRAD51-independent recombination by one-ended strand invasion coupled to DNA synthesis, followed by single-strand annealing. The Rad1/Rad10 endonuclease is required to trim intermediates formed during single-strand annealing and thus was expected to be required forRAD51-independent events by this model. Double-strand break repair between plasmid-borne inverted repeats was less efficient inrad1 rad51 double mutants than in rad1 andrad51 strains. In addition, repair events were delayed and frequently associated with plasmid loss. Furthermore, the repair products recovered from the rad1 rad51 strain were primarily in the crossover configuration, inconsistent with conservative models for mitotic double-strand break repair.

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A novel protein, Rsf1/Pxd1, is critical for the single-strand annealing pathway of double-strand break repair inSchizosaccharomyces pombe

Molecular Microbiology ◽

10.1111/mmi.13001 ◽

2015 ◽

Vol 96 (6) ◽

pp. 1211-1225 ◽

Cited By ~ 1

Author(s):

Hanqian Wang ◽

Zhanlu Zhang ◽

Lan Zhang ◽

Qiuxue Zhang ◽

Liang Zhang ◽

...

Keyword(s):

Strand Break ◽

Double Strand Break ◽

Single Strand ◽

Double Strand Break Repair ◽

Single Strand Annealing ◽

Break Repair ◽

Strand Annealing ◽

Novel Protein

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Analysis of DNA Double-Strand Break End Resection and Single-Strand Annealing in S. pombe

Homologous Recombination - Methods in Molecular Biology ◽

10.1007/978-1-0716-0644-5_4 ◽

2020 ◽

pp. 47-57

Author(s):

Zhenxin Yan ◽

Sandeep Kumar ◽

Grzegorz Ira

Keyword(s):

Strand Break ◽

Double Strand Break ◽

Single Strand ◽

Dna Double Strand Break ◽

Single Strand Annealing ◽

Strand Annealing ◽

End Resection

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mus309 mutation, defective in DNA double-strand break repair, affects intergenic but not intragenic meiotic recombination in Drosophila melanogaster

Genetics Research ◽

10.1017/s0016672305007883 ◽

2005 ◽

Vol 86 (3) ◽

pp. 185-191 ◽

Cited By ~ 10

Author(s):

PETTER PORTIN

Keyword(s):

X Chromosome ◽

Meiotic Recombination ◽

Strand Break ◽

Double Strand Break ◽

Double Strand Break Repair ◽

Crossing Over ◽

Dna Double Strand Break ◽

D Loop ◽

Break Repair ◽

Strand Annealing

The effect was investigated of the hypomorphic DNA double-strand break repair, notably synthesis-dependent strand annealing, deficient mutation mus309 on the third chromosome of Drosophila melanogaster on intergenic and intragenic meiotic recombination in the X chromosome. The results showed that the mutation significantly increases the frequency of intergenic crossing over in two of three gene intervals of the X chromosome studied. Interestingly the increase was most prevalent in the tip of the X chromosome where crossovers normally are least frequent per physical map unit length. In particular crossing over interference was also affected, indicating that the effect of the mus309 mutation involves preconditions of crossing over but not the event of crossing over itself. On the other hand, the results also show that most probably the mutation does not have any effect on intragenic recombination, i.e. gene conversion. These results are fully consistent with the present molecular models of meiotic crossing over initiated by double-strand breaks of DNA followed by formation of a single-end-invasion intermediate, or D-loop, which is subsequently processed to generate either crossover or non-crossover products involving formation of a double Holliday junction. In particular the results suggest that the mus309 gene is involved in resolution of the D-loop, thereby affecting the choice between double-strand-break repair (DSBR) and synthesis-dependent strand annealing (SDSA) pathways of meiotic recombination.

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Intermolecular recombination between DNAs introduced into mouse L cells is mediated by a nonconservative pathway that leads to crossover products.

Molecular and Cellular Biology ◽

10.1128/mcb.10.1.103 ◽

1990 ◽

Vol 10 (1) ◽

pp. 103-112 ◽

Cited By ~ 48

Author(s):

F L Lin ◽

K Sperle ◽

N Sternberg

Keyword(s):

Mammalian Cells ◽

Dna Recombination ◽

Strand Break ◽

Double Strand Break ◽

Yeast Cells ◽

Double Strand Break Repair ◽

L Cells ◽

Single Strand Annealing ◽

Break Repair ◽

Strand Annealing

We describe experiments designed to measure the efficiency of intermolecular recombination between mutant herpesvirus thymidine kinase (tk) genes introduced into mouse L cells. Recombinants were scored as stable transformants containing a functional tk gene. The two recombination substrates used were ptkB8, a pBR322-based plasmid containing a mutant tk gene, with a BamHI linker in an SphI restriction site that is centrally located within the gene, and mp10tk delta 3' delta 5', an mp10 vector with a tk gene deleted at both the 3' and 5' ends. The only homology shared by the two DNAs is 885 base pairs within the tk gene. To determine whether the double-strand break repair model that has been used to explain recombination in yeast cells (J. W. Szostak, T. L. Orr-Weaver, R. J. Rothstein, and F. W. Stahl, Cell 33:25-35, 1983) can account for recombination during the introduction of these DNAs into mammalian cells, we transformed cells with BamHI-linearized ptkB8 and supercoiled mp10tk delta 3' delta 5' replicative-form DNA. These two DNAs should recombine efficiently according to that model and should generate gene conversion products. In this reaction, the supercoiled DNA acts as the donor of information to repair the cleaved tk gene. Our results indicated that the efficiency of this reaction was very low (less than 10 transformants were obtained per 0.1 microgram of each DNA used in the reaction per 10(6) cells). In contrast, if BamHI-cleaved ptkB8 DNA was cotransformed into cells along with a circular DNA molecule containing a tk gene deleted only at its 3' end or only at its 5' end (mp10tk delta 3' or mp10tk delta 5'), then the efficiency of recombination could be more than 4 orders of magnitude higher than it was with circular mp10tk delta 3' delta 5' DNA. Recombination frequencies were highest when the tk delta 3' or tk delta 5' DNA used was cleaved at the tk deletion junction. Southern analyses of DNA from TK+ transformants generated with BamHI-cleaved ptkB8 and BamHI-cleaved mp10tk delta 3' DNAs indicated that recombination was almost always associated with the reassortment of markers flanking the reconstructed tk DNA. Together, these results are more consistent with the nonconservative single-strand annealing model for recombination that we proposed several years ago (F.-L. Lin, K. Sperle, and N. Sternberg, Mol. Cell. Biol. 4:1020-1034, 1984) than they are with the double-strand break repair model.

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