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 expression 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 cytoplasm of their eukaryotic host's cells upon contact with these cells. The secreted YopN protein (also known as LcrE) is required to block Yop secretion in the presence of calcium in vitro or before contact with a eukaryotic cell in vivo. In this study, we characterize the role of the tyeA, sycN and yscB gene products in the regulation of Yop secretion in Yersinia pestis. Mutants specifically defective in the expression of TyeA, SycN or YscB were no longer able to block Yop secretion in the presence of calcium. In addition, the secretion of YopN was specifically reduced in both the sycN and the yscB deletion mutants. Protein cross-linking and immunoprecipitation studies in conjunction with yeast two-hybrid analyses showed that SycN and YscB 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 alone, specifically associates with YopN. SycN and YscB share amino acid sequence 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.
Day
J B
JB
Department of Microbiology and Immunology, University of Miami School of Medicine, FL 33101, USA.
Plano
G V
GV
D000942:Antigens, Bacterial / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D016296:Mutagenesis; D010641:Phenotype; D052899:Pore Forming Cytotoxic Proteins; D011550:Pseudomonas aeruginosa; D015534:Trans-Activators / Q000235:genetics; D015008:Yersinia enterocolitica; D015010:Yersinia pestis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D015011:Yersinia pseudotuberculosis
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; C043908:LcrV protein, Yersinia; D008565:Membrane Proteins; D018832:Molecular Chaperones; D052899:Pore Forming Cytotoxic Proteins; C081269:SycE protein, Yersinia; C090049:SycH protein, Yersinia; C117910:SycN protein, Yersinia enterocolitica; D015534:Trans-Activators; C112196:TyeA protein, Yersinia; C117911:YscB protein, Yersinia pestis; C060772:yopM protein, Yersinia; C070015:YopN protein, Yersinia