A complex composed of SycN and YscB functions as a specific chaperone for YopN in Yersinia pestis

Human pathogenic Yersinia resist host defences, in part through the express ion and delivery of a set of plasmid-encoded virulence proteins termed Yops . A number of these Yops are exported from the bacteria directly into the c ytoplasm of their eukaryotic host's cells upon contact with these cells. Th e secreted YopN protein (also known as LcrE) is required to block Yop secre tion in the presence of calcium in vitro or before contact with a eukaryoti c cell in vivo. In this study, we characterize the role of the tyeA, sycN a nd yscB gene products in the regulation of Yop secretion in Yersinia pestis . Mutants specifically defective in the expression of TyeA, SycN or YscB we re no longer able to block Yop secretion in the presence of calcium. in add ition, the secretion of YopN was specifically reduced in both the sycH and the yscB deletion mutants. Protein crosslinking and immunoprecipitation stu dies in conjunction with yeast two-hybrid analyses showed that SycN and Ysc B interact with one another to form a SycN/YscB complex. Yeast three-hybrid analyses demonstrated that the SycN/YscB complex, but not SycN or YscB ato ne, specifically associates with YopN. SycN and YscB share amino acid seque nce similarity and structural similarities with the specific Yop chaperones SycE and SycH. Together, these results indicate that a complex composed of SycN and YscB functions as a specific chaperone for YopN in Y. pestis.