Mapping DNA damage‐dependent genetic interactions in yeast via party mating and barcode fusion genetics |
| |
| Authors: | J Javier Díaz‐Mejía Albi Celaj Joseph C Mellor Atina Coté Attila Balint Brandon Ho Pritpal Bansal Fatemeh Shaeri Marinella Gebbia Jochen Weile Marta Verby Anna Karkhanina YiFan Zhang Cassandra Wong Justin Rich D'Arcy Prendergast Gaurav Gupta Sedide Öztürk Daniel Durocher Grant W Brown Frederick P Roth |
| |
| Institution: | 1. Donnelly Centre, University of Toronto, Toronto, ON, Canada;2. Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada;3. Lunenfeld‐Tanenbaum Research Institute, Mt. Sinai Hospital, Toronto, ON, Canada;4. Department of Computer Science, University of Toronto, Toronto, ON, Canada;5. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA;6. Department of Biochemistry, University of Toronto, Toronto, ON, Canada;7. Center for Cancer Systems Biology (CCSB) and Department of Cancer Biology, Dana‐Farber Cancer Institute, Boston, MA, USA;8. Canadian Institute for Advanced Research, Toronto, ON, Canada |
| |
| Abstract: | Condition‐dependent genetic interactions can reveal functional relationships between genes that are not evident under standard culture conditions. State‐of‐the‐art yeast genetic interaction mapping, which relies on robotic manipulation of arrays of double‐mutant strains, does not scale readily to multi‐condition studies. Here, we describe barcode fusion genetics to map genetic interactions (BFG‐GI), by which double‐mutant strains generated via en masse “party” mating can also be monitored en masse for growth to detect genetic interactions. By using site‐specific recombination to fuse two DNA barcodes, each representing a specific gene deletion, BFG‐GI enables multiplexed quantitative tracking of double mutants via next‐generation sequencing. We applied BFG‐GI to a matrix of DNA repair genes under nine different conditions, including methyl methanesulfonate (MMS), 4‐nitroquinoline 1‐oxide (4NQO), bleomycin, zeocin, and three other DNA‐damaging environments. BFG‐GI recapitulated known genetic interactions and yielded new condition‐dependent genetic interactions. We validated and further explored a subnetwork of condition‐dependent genetic interactions involving MAG1, SLX4, and genes encoding the Shu complex, and inferred that loss of the Shu complex leads to an increase in the activation of the checkpoint protein kinase Rad53. |
| |
| Keywords: | condition‐dependent DNA barcode
en masse
genetic interaction sequencing |
|
|
