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Fig. 5 | BMC Microbiology

Fig. 5

From: HrcU and HrpP are pathogenicity factors in the fire blight pathogen Erwinia amylovora required for the type III secretion of DspA/E

Fig. 5

DspE secretion in Ea1189 strains. a Composite image of in vitro secretion of DspE-cyaA fusion proteins in minimal medium was visualized via western blot assay using an anti-CyaA antibody. DspE localization was detected in both the culture supernatant (S) and the cell pellet (P). Native HrcU, HrpP and the HrcUNPTH domain were required for DspE secretion as Ea1189∆hrcU/pRRM2 expressing HrcUN266A was unable to complement DspE secretion into the supernatant. HrcU and HrpP did not affect the production of DspE in the cell pellet. b Separate images composing (a). c T3S secretome in Ea1189 strains. All strains were cultured in minimal medium and after processing, were separated via one-dimensional SDS-PAGE and stained with silver nitrate. DspE is represented by a band corresponding to ~200 kDa and marked with an arrow. Both WT Ea1189 and Ea1189ΔhrcU/pRRM1 secrete DspE in vitro. Ea1189ΔhrcU and Ea1189ΔhrcU/pRRM2 cannot secret DspE indicating that HrcUN266A is required for DspE secretion. WT Ea1189 producing HrcUN266A does not exhibit a dominant negative effect on DspE secretion

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