Effect of SopE on stability of advanced Salmonella-containing vacuole (SCV)

Yiwei Weng

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Journal of Microbes and Infections ›› 2022, Vol. 17 ›› Issue (5) : 297-304.

Yiwei Weng

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Abstract

To study the effect of SopE, a type III secretory system effector encoded by Salmonella enterica serovar Typhi, on SCV induced by S. Typhi after invasion of macrophages. S.Typhi GIFU1007was used as the wild type (WT) strain to construct the sopE mutant (ΔsopE) by homologous recombination using the suicide plasmid pGMB151. The ΔsopE complementary strain (C-ΔsopE) was constructed with the plasmid complementary method by using pBAD33.The recombinant plasmid pET28-SFGFP was introduced into the WT, ΔsopE and C-ΔsopE , respectively, to yield WT/pBAD33::pET28-SFGFP, ΔsopE/pBAD33::pET28-SFGFP and C-ΔsopE/pET28-SFGFP. The effect of SopE on the intracellular viability of S. Typhi was studied by THP-1 intracellular viability assay of macrophages. The effect of SopE on the stability of late SCV was investigated by immunofluorescence and real-time quantitative PCR (qRT-PCR). The relationship between SopE and autophagy was further investigated by qRT-PCR and Western blot. The intracellular viability of THP-1 macrophages with ΔsopE/pBAD33::PET28-SFGFP was significantly lower than that of WT/pBAD33::PET28-SFGFP and C-ΔsopE/PET28-SFGFP. The immunofluorescence assay and qRT-PCR results showed that deletion of sopE decreased the stability of late SCV. The qRT-PCR and Western blot results showed that SopE inhibited the expression of autophagy related genes. The data presented here showed that SopE regulated the stability of late SCV by affecting autophagy.

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Yiwei Weng. Effect of SopE on stability of advanced Salmonella-containing vacuole (SCV)[J]. Journal of Microbes and Infections. 2022, 17(5): 297-304

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