Abstract
Type 3 secretion systems form an integral part of the arsenal of many pathogenic bacteria. These injection machines, together with their cargo of subversive effector proteins, are capable of manipulating the cellular environment of the host in order to ensure persistence of the pathogen. In order to fully appreciate the functions of Type 3 effectors, it is necessary to gain spatio‐temporal knowledge of each effector during the process of infection. A number of genetic modifications have been exploited in order to reveal effector protein secretion, translocation and subsequent activity, and localisation within host cells. In this review, we will discuss the many available approaches for tracking effector protein dynamics and discuss the challenges faced to improve the current technologies and gain a clearer picture of effector protein function.
Highlights
The successful proliferation of a bacterial species depends upon their ability to colonise an appropriate niche
The Type 3 secretion systems (T3SS) is a 3.5 megadalton complex consisting of a number of bacterial membrane embedded components, a hollow proteinaceous needle, and a tip translocon apparatus, which becomes embedded in the host cell membrane thereby allowing the formation of a continuous conduit from the bacterial cytosol to the host cytosol (Marlovits et al, 2004; Radics, Königsmaier, & Marlovits, 2014)
To improve expression and resultant fluorescence intensity from bacterial expression constructs, codon usage of the iLOV domain was optimised for E. coli to produce phiLOV2.1 (Gawthorne et al, 2016). Fusion of this optimised LOV domain to Escherichia coli (EHEC) translocated intimin receptor (Tir) did not interfere with effector translocation or functionality, and Tir‐phiLOV could be readily visualised both in bacterial cells prior to secretion and within A/E lesions on the host cell after bacterial docking (Gawthorne et al, 2016)
Summary
The successful proliferation of a bacterial species depends upon their ability to colonise an appropriate niche. Translocation of effector proteins can be determined microscopically using enzymatic tags that alter the fluorescence spectrum of a substrate in the host cell cytosol.
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