The Site-Specific Recombination System of the<i>Escherichia coli</i>Bacteriophage φ24<sub>B</sub>

Mohaisen, Mohammed Radhi, McCarthy, Alan John, Adriaenssens, Evelien M ORCID: 0000-0003-4826-5406 and Allison, Heather Elizabeth ORCID: 0000-0003-0017-7992
(2020) The Site-Specific Recombination System of the<i>Escherichia coli</i>Bacteriophage φ24<sub>B</sub>. FRONTIERS IN MICROBIOLOGY, 11. 578056-.

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Stx bacteriophages are members of the lambdoid group of phages and are responsible for Shiga toxin (Stx) production and the dissemination of Shiga toxin genes (<i>stx</i>) across shigatoxigenic <i>Escherichia coli</i> (STEC). These toxigenic bacteriophage hosts can cause life-threatening illnesses, and Stx is the virulence determinant responsible for the severe nature of infection with enterohemorrhagic <i>E. coli</i>, a subset of pathogenic STEC. Stx phages are temperate, and in the present study, the identification of what is actually required for Stx phage Φ24<sub>B</sub> and bacterial DNA recombination was tested using both <i>in vitro</i> and <i>in situ</i> recombination assays. It is well established that phage λ, which underpins most of what we understand about lambdoid phage biology, requires its own encoded phage attachment site (<i>attP</i>) of 250 bp, a host-encoded attachment site (<i>attB</i>) of 21 bp, and a host-encoded DNA binding protein known as integration host factor (IHF). The assays applied in this study enabled the manipulation of the phage attachment site (<i>attP</i>) and the bacterial attachment site (<i>attB</i>) sequences and the inclusion or exclusion of a host-encoded accessory element known as integration host factor. We were able to demonstrate that the minimal <i>attP</i> sequence required by Φ24<sub>B</sub> phage is between 350 and 427 bp. Unlike phage λ, the minimal necessary flanking sequences for the <i>attB</i> site do not appear to be equal in size, with a total length between 62 and 93 bp. Furthermore, we identified that the Φ24<sub>B</sub> integrase does not require IHF to drive the integration and the recombination process. Understanding how this unusual Stx phage integrase works may enable exploitation of its promiscuous nature in the context of genetic engineering.

Item Type: Article
Uncontrolled Keywords: Shiga toxin, site-specific DNA recombination, Stx phage, prophage, prophage integrase, tyrosine recombinase, attP, attB
Depositing User: Symplectic Admin
Date Deposited: 15 Sep 2020 08:54
Last Modified: 05 Oct 2023 20:53
DOI: 10.3389/fmicb.2020.578056
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