Factors affecting template switch recombination associated with restarted DNA replication
- University of Oxford, United Kingdom
Figures
Figure 1 with 1 supplement
TS downstream of RTS1.
(A) Map showing sites of insertion for the direct repeat recombination reporter on chromosome 3. (B) Schematic of the direct repeat recombination reporter showing the position of RTS1 insertion and …
Figure 1—figure supplement 1
Effect of ori1253∆ on the frequency of TS recombination 75 kb downstream of RTS1.
Ade+ recombinant frequencies for strains MCW7430, MCW7987, MCW7616 and MCW7620. Data are mean values ± SD. Ade+ recombinant frequencies with statistical analysis are also shown in Supplementary file …
Figure 2 with 1 supplement
Effect of pfh1-m21 and pfh1-mt* mutations on TS recombination at and downstream of RTS1.
(A) Ade+ recombinant frequencies for strains MCW4712, MCW4940, MCW4954, MCW7131, MCW7599, MCW7603, MCW7257, MCW7421 and MCW7425. (B) Ade+ recombinant frequencies for strains MCW4713, MCW4942, …
Figure 2—figure supplement 1
Relative Ade+ recombinant frequencies for the pfh1-m21 and pfh1-mt* strains shown in Figure 2B.
Values are relative to wild-type.
Figure 3
Pfh1 is needed for replication past RTS1-AO.
(A) Schematic showing location of RTS1-AO and hphMX6 adjacent to bub1 on chromosome 3. The position of the probe used for the 2DGE analysis in B) is also shown. (B) 2DGE of replication intermediates …
Figure 4 with 1 supplement
Effect of ori1253∆ on the frequency of TS recombination downstream of RTS1 in wild-type and pfh1 mutants.
The strains are MCW7414, MCW7598, MCW7602, MCW7293, MCW7423, MCW7427, MCW7416, MCW7600, MCW7604, MCW7295, MCW7424, MCW7428. Data are mean values ± SD. Ade+ recombinant frequencies with statistical …
Figure 4—figure supplement 1
Relative Ade+ recombinant frequencies for the pfh1-m21 and pfh1-mt* strains shown in Figure 4.
Values are relative to wild-type.
Figure 5
Effect of tRNAGLU08CD and tRNAGLU08HO on TS downstream of RTS1.
(A) Schematic showing the position of tRNAGLU08CD/HO within the direct repeat recombination reporter 12.4 kb downstream of RTS1. (B) Ade+ recombinant frequencies for strains MCW7229, MCW7434 and …
Figure 6 with 2 supplements
Effect of rqh1∆, srs2∆, fbh1∆, fml1∆, mus81∆, pcf1∆ and pcf2∆ on TS recombination at and downstream of RTS1.
(A) Ade+ recombinant frequencies for strains MCW4712, MCW1443, FO1748, FO1814, MCW3059, MCW1451, MCW6972 and MCW7147. (B) Ade+ recombinant frequencies for strains MCW4713, MCW1447, FO1750, FO1816, …
Figure 6—figure supplement 1
Relative Ade+ recombinant frequencies for the RTS1-AO containing strains shown in Figure 5.
Values are relative to wild-type.
Figure 6—figure supplement 2
Fml1 and Mus81 are not needed for replication past RTS1-AO.
(A) Schematic showing location of RTS1-AO between ade6 heteroalleles on chromosome 3. The position of the probe used for the 2DGE analysis in (B) is also shown. (B) 2DGE of replication intermediates …
Figure 7
Model for TS between direct repeats.
Parental DNA strands are dark blue, nascent DNA strands are light blue and DNA repeats are yellow. The grey line in steps 5, 6, 7a and 8a indicates that the light blue and yellow lines, which it …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (S. pombe) | various strains | PMID: 15889146 | standard laboratory strain (972) derivatives; see Supplementary file 2 | |
| Strain, strain background (S. pombe) | various strains | PMID: 25806683 | standard laboratory strain (972) derivatives; see Supplementary file 2 | |
| Strain, strain background (S. pombe) | various strains | this paper | standard laboratory strain (972) derivatives; see Supplementary file 2 | |
| Strain, strain background (S. pombe) | MCW4956 | PMID: 22426535 | standard laboratory strain (972) derivatives; see Supplementary file 2 | |
| Strain, strain background (S. pombe) | various strains | PMID: 19546232 | standard laboratory strain (972) derivatives; see Supplementary file 2 | |
| Strain, strain background (S. pombe) | MCW3059 | PMID: 18851838 | standard laboratory strain (972) derivatives; see Supplementary file 2 | |
| Strain, strain background (S. pombe) | MCW3061 | PMID: 18851838 | standard laboratory strain (972) derivatives; see Supplementary file 2 | |
| Recombinant DNA reagent | pMJ33 | this paper | plasmid; see Materials and methods | |
| Recombinant DNA reagent | pMJ34 | this paper | plasmid; see Materials and methods | |
| Recombinant DNA reagent | pCB44 | this paper | plasmid; see Materials and methods | |
| Recombinant DNA reagent | pMW899 | PMID: 22426535 | plasmid | |
| Recombinant DNA reagent | pMW905 | PMID: 22426535 | plasmid | |
| Sequence-based reagent | various oligonucleotides | this paper | see Supplementary file 3 (oligonucleotides) |
Additional files
-
Supplementary file 1
Direct repeat recombinant frequencies.
- https://doi.org/10.7554/eLife.41697.014
-
Supplementary file 2
Schizosaccharomyces pombe strains.
- https://doi.org/10.7554/eLife.41697.015
-
Supplementary file 3
Oligonucleotides.
- https://doi.org/10.7554/eLife.41697.016
-
Transparent reporting form
- https://doi.org/10.7554/eLife.41697.017
