Key Resources Table
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit polyclonal anti-RADX | Novus Biologicals | Cat#NBP2-13887; RRID: AB_2687552 |
| Mouse monoclonal anti-RAD51 (14B4) | Abcam | Cat# ab213, RRID:AB_302856 |
| Mouse monoclonal anti FLAG M2 | Sigma Aldrich | Cat#F3165; RRID:AB_259529 |
| Rabbit polyclonal anti-GFP | Santa Cruz | Cat#sc-8334; RRID:AB_641123 |
| Rabbit polyclonal to GST | Abcam | Cat# ab111947 AB_10861266 |
| Mouse monoclonal anti-phospho-Histone H2A.X (Ser139) | Millipore | Cat#16-193, RRID:AB_310795 |
| Mouse monoclonal anti BrdU | BD | Cat#347580; RRID:AB_10015219 |
| Rat monoclonal anti BrdU | Abcam | Cat#ab6326; RRID:AB_305426 |
| Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 | Life Technologies | Cat# A32723; RRID:AB_2633275 |
| Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 | Life Technologies | Cat# A11007; RRID:AB_10561522 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| CldU | Sigma Aldrich | Cat#C6891 |
| IdU | Sigma Aldrich | Cat#l7125 |
| EdU | VICB synthesis core | N/A |
| Alexa Fluor 647 azide | Thermo Fisher | Cat#A10277 |
| Hydroxyurea | Sigma Aldrich | Cat#H8627 |
| MBP-RADX | This study | N/A |
| MBP-RADX OB2m | This study | N/A |
| MBP-RADX QVPK | This study | N/A |
| GFP-RADX | This study | N/A |
| RAD51 | Provided by M.M. | N/A |
| RAD51 K133R | Provided by M.M. | N/A |
| RAD51 K133A | Provided by M.M. | N/A |
| RPA | Provided by W.J.C. | N/A |
| GST-BRC3/4 | Provided by Patrick Sung | N/A |
| RPA-mCherry | Provided by E.J.C. | N/A |
| Critical Commercial Assays | ||
| Duolink In Situ PLA Probe Anti-Rabbit PLUS | Sigma Aldrich | Cat#DU092002 |
| Duolink In Situ PLA Probe Anti-Rabbit MINUS | Sigma Aldrich | Cat#DU092004 |
| Duolink In Situ Detection Reagents, Red | Sigma Aldrich | Cat#DU092008 |
| Coverslips | MicroSurfaces Inc. | N/A |
| Experimental Models: Cell Lines | ||
| U2OS | ATCC | Cat#HTB-96; RRID:CVCL_0042 |
| HEK293T | ATCC | Cat#CRL-11268; RRID:CVCL_1926 |
| U2OS ΔRADX | (Dungrawala et al., 2017) | N/A |
| Oligonucleotides | ||
| dT60: 5’-TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT-3’ | IDT | N/A |
| dT72: 5’-TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT-3′ | IDT | N/A |
| OligoD1: 5’-AAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTT-3’ | IDT | N/A |
| Recombinant DNA | ||
| Plasmid pDC1272 = pLEFP-FLAG-RADX | This study | N/A |
| Plasmid pMA32 = pLEGFP FLAG-RADX QVPK | This study | N/A |
| Plasmid pMA33 = pLEGFP-FLAG-RADX IV | This study | N/A |
| Plasmid pMA34 = pLEGFP-FLAG-RADX LLY | This study | N/A |
| Plasmid pMA35 = pLEGFP-FLAG-RADX R389E | This study | N/A |
| Plasmid pMA36 = pLEGFP-FLAG-RADX LF | This study | N/A |
| Plasmid pMA37 = pLEGFP-FLAG-RADX ELDD | This study | N/A |
| Plasmid pKB06 = pEF1a-RADX | This study | N/A |
| Plasmid pKB29= pEF1a-RADX OB2m | This study | N/A |
| Plasmid pMA39= pEF1a-RADX-QVPK | This study | N/A |
| Software and Algorithms | ||
| GraphPad Prism8/9 | GraphPad Software | https://www.graphpad.com/scientific-software/prism/; RRID: SCR_000306 |
| ImageLab | BioRad | http://www.biorad.com/en-us/sku/1709690-image-lab-software; RRID:SCR_014210 |
| MODELLER v9.16 | (Eswar et al., 2006) | https://salilab.org/modeller; RRID:SCR_008395 |
| PyMOL Version 2.0 | The PyMOL Molecular Graphics System, Version 2.0 Schrödinger, LLC. | https://www.pymol.org/; RRID:SCR_000305 |
| Scipion | (de la Rosa-Trevin et al., 2016) | http://scipion.i2pc.es; RRID:SCR_016738 |
| Deposited Data | ||
| Unprocessed blots, gels and microscopy images | This paper | https://dx.doi.org/10.17632/g3rfkjw225.1 |