ATPase-powered antiphage nuclease with tRNA stem-severing activity

This work aims to solve the age-old mystery of the physiological target of the OLD (overcoming lysogenization defect) nuclease of phage P2, which interferes with the replication of phage Lambda in P2 lysogens. We screened a library of 245 unique phage-encoded tRNAs in E. coli MG1655 engineered to recapitulate the growth defect induced by P2-OLD. To identify rescue tRNAs, next-generation sequencing was run on cells transformed with the phage tRNA library before ("input") and after activation of P2-OLD ("output").