High-throughput discovery of phage receptors using transposon insertion sequencing of bacteria.

Proceedings of the National Academy of Sciences of the United States of America
Kaitlyn E KortrightPaul E Turner

Abstract

As the most abundant microbes on Earth, novel bacteriophages (phages; bacteria-specific viruses) are readily isolated from environmental samples. However, it remains challenging to characterize phage-bacteria interactions, such as the host receptor(s) phages bind to initiate infection. Here, we tested whether transposon insertion sequencing (INSeq) could be used to identify bacterial genes involved in phage binding. As proof of concept, results showed that INSeq screens successfully identified genes encoding known receptors for previously characterized viruses of Escherichia coli (phages T6, T2, T4, and T7). INSeq screens were then used to identify genes involved during infection of six newly isolated coliphages. Results showed that candidate receptors could be successfully identified for the majority (five of six) of the phages; furthermore, genes encoding the phage receptor(s) were the top hit(s) in the analyses of the successful screens. INSeq screens provide a generally useful method for high-throughput discovery of phage receptors. We discuss limitations of our approach when examining uncharacterized phages, as well as usefulness of the method for exploring the evolution of broad versus narrow use of cellular receptors amo...Continue Reading

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Citations

Oct 14, 2020·PLoS Biology·Vivek K MutalikAdam P Arkin
Jan 13, 2021·Pediatric Pulmonology·Benjamin K ChanPaul E Turner
Jan 17, 2021·International Journal of Molecular Sciences·Anna Allué-GuardiaJordi B Torrelles
Nov 18, 2020·Current Opinion in Biotechnology·Fernando L Gordillo Altamirano, Jeremy J Barr
Apr 12, 2021·Biochemistry. Biokhimii︠a︡·Artem B IsaevKonstantin V Severinov
May 15, 2021·Environmental Microbiology·Castillo DanielMiddelboe Mathias
Jun 5, 2021·Proceedings of the National Academy of Sciences of the United States of America·Joshua M BorinJustin R Meyer
Jul 6, 2021·Frontiers in Microbiology·Kaitlyn E KortrightPaul E Turner
Aug 28, 2021·Microorganisms·Shazeeda KoonjanAnders S Nilsson
Sep 25, 2021·FEMS Microbiology Reviews·Julia E EgidoStan J J Brouns
Oct 13, 2021·PLoS Biology·Erin L AttrillStefano Pagliara
Oct 7, 2020·Current Biology : CB·Alita R Burmeister, Paul E Turner
Nov 5, 2021·Nature Reviews. Microbiology·Jiawei SunKerwyn Casey Huang

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Methods Mentioned

BETA
bacterial knockout
INSeq

Software Mentioned

Python scripts
EcoCyc
INSeq
Bowtie

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