Preprint Article Version 1 This version is not peer-reviewed

Genome-wide Screens Reveal Escherichia Coli Genes Required for Growth of T1-like Phage LL5 and rV5-like Phage LL12

Version 1 : Received: 17 September 2018 / Approved: 18 September 2018 / Online: 18 September 2018 (10:00:35 CEST)

How to cite: Piya, D.; Lessor, L.; Koehler, B.; Stonecipher, A.; Cahill, J.; Gill, J.J. Genome-wide Screens Reveal Escherichia Coli Genes Required for Growth of T1-like Phage LL5 and rV5-like Phage LL12. Preprints 2018, 2018090340 (doi: 10.20944/preprints201809.0340.v1). Piya, D.; Lessor, L.; Koehler, B.; Stonecipher, A.; Cahill, J.; Gill, J.J. Genome-wide Screens Reveal Escherichia Coli Genes Required for Growth of T1-like Phage LL5 and rV5-like Phage LL12. Preprints 2018, 2018090340 (doi: 10.20944/preprints201809.0340.v1).

Abstract

Factors affecting the host-virus interaction must be understood for the effective application of bacteriophages to combat bacterial pathogens. Two novel E. coli phages, the T1-like siphophage LL5 and the rV5-like myophage LL12, were subjected to forward genetic screens against the Keio collection, a library of single non-essential gene deletions in E.coli str. BW25113. These genome-wide screens and subsequent experiments identified eight genes required for efficient propagation of phage LL5 and six genes required for propagation of LL12. The majority of the genes identified were involved in production of the phage receptors. E. coli mutants deficient in heptose II and the phosphoryl substituent of heptose I of the inner core lipopolysaccharide (LPS) were unable to propagate phage LL5, as were mutants deficient in the outer membrane protein TolC. Mutants lacking glucose I of the LPS outer core failed to propagate LL12. Two cytoplasmic chaperones, PpiB and SecB, were found to be required for efficient propagation of phage LL5 but not LL12. This approach may be useful for identifying phage receptors and required host factors in other phages, which would provide valuable information for their potential use as therapeutics and for phage engineering.

Subject Areas

bacteriophage; phage-host interactions; phage receptors; high-throughput screen; tail fibers

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