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Mapping DNA damage-dependent genetic interactions in yeast via party mating and barcode fusion genetics

By 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

Posted 31 Aug 2017
bioRxiv DOI: 10.1101/181750 (published DOI: 10.15252/msb.20177985)

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 and 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 a decrease in the activation or activity of the checkpoint protein kinase Rad53.

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