1887

Abstract

is an important Gram-negative enteric pathogen affecting both animals and humans. It possesses a type III secretion system (T3SS) essential for pathogenesis. EseB, EseC and EseD have been shown to form a translocon complex after secretion, while EscC functions as a T3SS chaperone for EseB and EseD. In this paper we identify EscA, a protein required for accumulation and proper secretion of another translocon component, EseC. The gene is located upstream of and the EscA protein has the characteristics of T3SS chaperones. Cell fractionation experiments indicated that EscA is located in the cytoplasm and on the cytoplasmic membrane. Mutation with in-frame deletion of greatly decreased the secretion of EseC, while complementation of restored the wild-type secretion phenotype. The stabilization and accumulation of EseC in the cytoplasm were also affected in the absence of EscA. Mutation of did not affect the transcription of but reduced the accumulation level of EseC as measured by using an EseC-LacZ fusion protein in . Co-purification and co-immunoprecipitation studies demonstrated a specific interaction between EscA and EseC. Further analysis showed that residues 31–137 of EseC are required for EseC-EscA interaction. Mutation of EseC residues 31–137 reduced the secretion and accumulation of EseC in . Finally, infection experiments showed that mutations of EscA and residues 31–137 of EseC increased the LD by approximately 10-fold in blue gourami fish. These results indicated that EscA functions as a specific chaperone for EseC and contributes to the virulence of .

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2009-04-01
2019-11-12
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