Tracking the dynamic interplay between bacterial and host factors during pathogen-induced vacuole rupture in real time.
Cell Microbiol. 2010 Apr 1; 12(4):545-56
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I found this article interesting because the authors developed a novel fluorescence microscopy methodology to detect rupture of the phagosome membrane by intracellular bacterial pathogens at the single cell level.
By combining the use of a chimeric protein AIDA-beta-lactamase, FRET, microscopy and an automated algorithm the authors were able to detect bacterial escape from phagosomes into the cytoplasm in real time. This novel procedure is temporally sensitive compared to other methods used to detect bacterial escape, like comet tail formation. Additionally, the use of fluorescent fusions to different host cell proteins helped them to identify multiple proteins recruited during the invasion and vacuole escape processes (i.e. in Shigella escape, RhoA, Rac1 and galectin-3 are recruited). Using this system, the authors were also able to show the role of Salmonella effectors PipB2 and SifA in vacuole stabilization. This method is limited so far to Gram-negative bacteria but could potentially be adapted for other pathogenic bacteria.
Steele-Mortimer O and Ibarra J: F1000Prime Recommendation of [Ray K et al., Cell Microbiol 2010, 12(4):545-56]. In F1000Prime, 13 Apr 2010; DOI: 10.3410/f.2886956.2555054. F1000Prime.com/2886956#eval2555054
F1000Prime Recommendations, Dissents and Comments for [Ray K et al., Cell Microbiol 2010, 12(4):545-56]. In F1000Prime, 03 Sep 2014; F1000Prime.com/2886956
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