Yersinia enterocolitica type III secretion. On the role of SycE in targeting YopE into HeLa cells.
Yersinia enterocolitica inject toxic proteins (effector Yops) into the cytosol of eukaryotic cells by a mechanism requiring the type III machinery. Previous work mapped a signal sufficient for the targeting of fused reporter proteins to amino acids 1-100 of YopE. Targeting requires the binding of SycE to YopE residues 15-100 in the bacterial cytoplasm. We asked whether SycE functions only to stabilize YopE in the bacterial cytoplasm, or whether the secretion chaperone itself contributes to substrate recognition by the type III machinery. Fusions of glutathione S-transferase to either the N or C terminus of SycE resulted in hybrid proteins that bound YopE but prevented targeting of the export substrate into HeLa cells. As compared with wild-type SycE, glutathione S-transferase-SycE bound and stabilized YopE in the bacterial cytoplasm but failed to release the polypeptide for export by the type III machinery. Thus, it appears that SycE functions to deliver YopE to the type III secretion machinery. A model is presented that accounts for substrate recognition of effector Yops, a group of proteins that do not share amino acid sequence or physical similarities.
Cheng
L W
LW
Department of Microbiology and Immunology, University of California Los Angeles School of Medicine, Los Angeles, California 90095, USA.
Schneewind
O
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D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D017931:DNA Primers; D005982:Glutathione Transferase / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D016133:Polymerase Chain Reaction; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D015008:Yersinia enterocolitica / Q000235:genetics / Q000502:physiology*
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; C081269:SycE protein, Yersinia; D015534:Trans-Activators; C063239:yopE protein, Yersinia; D005982:Glutathione Transferase