KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-Digoxigenin Fab Fragment (from Sheep) | Roche | Cat# 11214667 |
| Bacterial and Virus Strains | ||
| E. coli Bl21 Rosetta2 (DE3) F−ompThsdSB(rB− mB−) gal dcm (DE3) pRARE2 (CamR) | EMD Millipore | Cat# 714003 |
| E. coli NEB Turbo F proA+B+ lacIqΔ lacZ M15/ fhuA2 Δ(lac-proAB) glnV gal R(zgb-210::Tn10)TetSendA1 thi-1 Δ(hsdS-mcrB)5 | NEB | Cat# C2984H |
| E. coli DH5alpha fhuA2 a(argF-lacZ)U169 phoA glnV44 a80a(lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17 | NEB | Cat# C2987H |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Sc GST-Rad54 | This Study | N/A |
| Sc GST-GFP-Rad54 | This Study | N/A |
| Sc Rad51 | This Study | N/A |
| Sc RPA-6xHiS | This Study | N/A |
| Sc RPA-GFP-6xHis | This Study | N/A |
| Hs RPA-GFP-6xHis | This Study | N/A |
| Ec SSB-GFP-6xHis | This Study | N/A |
| Sc Nap1 | This Study | N/A |
| Sc Histone H2AS47C | This Study | N/A |
| Sc Histone H2B | This Study | N/A |
| Sc Histone H3 | This Study | N/A |
| Sc Histone H4 | This Study | N/A |
| Streptavidin from Streptomyces | Millipore | Cat# S4762 |
| Albumin from bovine serum | Sigma-Aldrich | Cat# A7030 |
| Sc Ulp1-SUMO Protease | Sigma-Aldrich | Cat# SAE0067 |
| Talon Resin | Clontech | Cat# 635503 |
| HisTrap prepacked column | GE Healthcare | Cat# 17-5247-01 |
| GST-Resin | GE Healthcare | Cat# 17-0756-01 |
| Hi-load 16/60 Superdex-200 | GE Healthcare | Cat# 17-1043-01 |
| Sephacryl-S-300 | GE Healthcase | Cat# 17-0599-10 |
| Q-Sepharose | GE Healthcare | Cat# 17-0510-01 |
| Dynabeads M-280 Streptavidin | Invitrogen | Cat# 11205D |
| 1,2-dioleoyl-sn-glycero-3-phosphocholine | Avanti Polar Lipids | Cat# 850375C |
| 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl) (sodium salt) | Avanti Polar Lipids | Cat# 870273P |
| 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(methoxy(polyethylene glycol)-2000](ammonium salt) | Avanti Polar Lipids | Cat# 880130P |
| Critical Commercial Assays | ||
| In-Fusion HD Cloning plus Kit | Clontech | Cat# 638909 |
| Deposited data | ||
| Kymographs used for all data analysis | Mendeley | https://doi.org/10.17632/x2cb2mkhty.1 |
| Experimental Models: Organisms/Strains | ||
| S. cerevisiae (MAT alpha leu2 trp1 ura3–52 prb1–1122 pep4–3 his3::pGAL1-GAL4) | Gift from Symington Lab (Columbia University Medical Center) | LSY0929 |
| S. cerevisiae rad51Δ (MATalpha leu2–3,112 trp1–1 can1–100ura3–1 ade2-1 his3–11,15 rad51:: URA3, rad5–535) | Gift from Symington Lab (Columbia University Medical Center) | LSY0411 |
| S. cerevisiae (MAT a, ura3–52 trp1 leu2Δ his3Δ200 pep4::HIS3 prb1D1.6R can, GAL) | Gift from Symington Lab (Columbia University Medical Center) | RDKY2275 |
| Oligonucleotides | ||
| Position 8.6 (21 -nt homology) 5′-AAT TCT CAT TTT ACT TAC CGG ACG CTA TTA GCA GTG GGT GAG CAA AAA CAG GAA GGC-3′ | IDT, This Study | N/A |
| Position 8.6 (15-nt homology) 5′-AAT TCT CAT TTT ACT TAC CGG ACG CTA TTA GCA GTG ATG ATC CAA AAA CAG GAA GGC-3′ | IDT, This Study | N/A |
| Position 8.6 (9-nt homology) 5′-AAT TCT CAT TTT ACT TAC CGG ACG CTA TTA GCA GTG ATG ATC CAT ACT CAG GAA GGC-3′ | IDT, This Study | N/A |
| Position 8.6 (0-nt homology) 5′-AAT TCT CAT TTT ACT TAC CGG ACG CTA TTA GCA GTG CAT AGT ATC CCT AGT CCG TAA-3′ | IDT, This Study | N/A |
| Position 8.6 90 nt homology 5-GGT GCA CGA GTG GGT TAC ATC GAA CTG GAT CTC AAC AGC GGT AAG ATC CTT GAG AGT TTT CGC CCC GAA GAA CGT TTT CC A ATG ATG AGC | IDT, This Study | N/A |
| Position 8.6 150 nt homology GTA AAA GAT GCT GAA GAT CAG TTG GGT GCA CGA GTG GGT TAC ATC GAA CTG GAT CTC AAC AGC GGT AAG ATC CTT GAG AGT TTT CGC CCC GAA GAA CGT TTT CCA ATG ATG AGC ACT TTT AAA GTT CTG CTA TGT GGC GCG GTA TTA TCC | IDT, This Study | N/A |
| Position 4.5 (21-nt homology) 5′-AAT TCT CAT TTT ACT TAC CGG ACG CTA TTA GCA GTG GCG CGC CTG TGC ACT CTG TGG-3′ | IDT, This Study | N/A |
| Labeled strand 5′-Atto565-CAC TGC TAA TAG CGT CCG GTA AGT AAA ATG AGA ATT-3′ | IDT, This Study | N/A |
| Labeled strand 5′-Atto647N-CAC TGC TAA TAG CGT CCG GTA AGT AAA ATG AGA ATT-3′ | IDT, This Study | N/A |
| For additional oligonucleotide sequences see Key Resource Table S1 | This Paper | N/A |
| Other | ||
| Fisherfinest Premium Cover Glass | Fisher Scientific | 12-548-5P |
| Quartz Microscope Slides (1” × 3”) | Finkenbeiner | N/A |
| TIRFM Microscope | This Study | N/A |
| Recombinant DNA | ||
| pUC19 plasmid | New England Biolabs | Cat# N3041S |
| CURMIDDNA | This Study | N/A |
| pYes2-GST-Rad54 | WD Heyer Lab (University of California, Davis) | N/A |
| pYes2-GFP-GST-Rad54 | This Study | N/A |
| pET28D-Nap1 | K. Luger Lab (University of Colorado, Boulder) | N/A |
| pET11C-H2A, H2B, H3, H4 | K. Luger Lab (University of Colorado, Boulder) | N/A |
| pET11C-SUMO-Rad51 | This study | N/A |
| p11D-sctRPA, sctRPA-GFP-70, sctRPA, hstRPA-GFP-70 | M. Wold Lab (University of Iowa) | N/A |
| Software and Algorithms | ||
| Graphpad Prism 8.2 | Graphpad | https://www.graphpad.com/scientific-software/prism/ |
| Nikon Elements Ars 5.11 | Nikon | (https://www.nikon.com/products/microscope-solutions/lineup/img_soft/nis-elements/table_of_features.htm |