A forward-genetic screen and dynamic analysis of lambda phage host-dependencies reveals an extensive interaction network and a new anti-viral strategy.

PLoS Genetics
Nathaniel D MaynardMarkus W Covert

Abstract

Latently infecting viruses are an important class of virus that plays a key role in viral evolution and human health. Here we report a genome-scale forward-genetics screen for host-dependencies of the latently-infecting bacteriophage lambda. This screen identified 57 Escherichia coli (E. coli) genes--over half of which have not been previously associated with infection--that when knocked out inhibited lambda phage's ability to replicate. Our results demonstrate a highly integrated network between lambda and its host, in striking contrast to the results from a similar screen using the lytic-only infecting T7 virus. We then measured the growth of E. coli under normal and infected conditions, using wild-type and knockout strains deficient in one of the identified host genes, and found that genes from the same pathway often exhibited similar growth dynamics. This observation, combined with further computational and experimental analysis, led us to identify a previously unannotated gene, yneJ, as a novel regulator of lamB gene expression. A surprising result of this work was the identification of two highly conserved pathways involved in tRNA thiolation-one pathway is required for efficient lambda replication, while the other has an...Continue Reading

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Citations

Aug 8, 2014·PLoS Computational Biology·Cheemeng TanLingchong You
Jul 3, 2013·Proceedings of the National Academy of Sciences of the United States of America·Xiaoqiu LiuJeffrey W Roberts
Jun 29, 2011·Proceedings of the National Academy of Sciences of the United States of America·Cesar O FloresJoshua S Weitz
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Methods Mentioned

BETA
gene knockout
PCR

Software Mentioned

MatLab

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