Yersinia enterocolitica type III secretion: evidence for the ability to transport proteins that are folded prior to secretion

BMC Microbiology
Gottfried WilharmJürgen Heesemann

Abstract

Pathogenic Yersinia species (Y. enterocolitica, Y. pestis, Y. pseudotuberculosis) share a type three secretion system (TTSS) which allows translocation of effector proteins (called Yops) into host cells. It is believed that proteins are delivered through a hollow needle with an inner diameter of 2-3 nm. Thus transport seems to require substrates which are essentially unfolded. Recent work from different groups suggests that the Yersinia TTSS cannot accommodate substrates which are folded prior to secretion. It was suggested that folding is prevented either by co-translational secretion or by the assistance of specific Yop chaperones (called Sycs). In this study we have fused YopE secretion signals of various length to the mouse dihydrofolate reductase (DHFR) in order to analyse the DHFR folding state prior to secretion. We could demonstrate that secretion-deficient as well as secretion-competent YopE-DHFR fusions complexed to SycE can be efficiently purified from Yersinia cytosol by affinity chromatography using methotrexate-agarose. This implies the folding of the DHFR fusion moiety despite SycE binding and contradicts the previously presented model of folding inhibition by chaperone binding. Secretion-deficient YopE-DHFR fusi...Continue Reading

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Citations

Oct 4, 2012·Journal of the American Chemical Society·Tracy C HolmesChaitan Khosla
Feb 27, 2009·The Journal of Biological Chemistry·Annika SchmidGottfried Wilharm
Oct 17, 2007·Nature Reviews. Microbiology·Armen Y MulkidjanianEugene V Koonin
Jul 8, 2005·The Journal of Biological Chemistry·Martin LocherGottfried Wilharm
Sep 6, 2012·Applied Microbiology and Biotechnology·Vic NorrisJason A Rosenzweig
Sep 5, 2015·The Journal of Biological Chemistry·Frédéric H Login, Hans Wolf-Watz
Nov 14, 2020·Biomolecules·Xiang-Yu Zhuang, Chien-Jung Lo
Feb 25, 2010·Microbes and Infection·Roland ArnoldThomas Rattei

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Methods Mentioned

BETA
electrophoresis
affinity purification
isothermal titration calorimetry
electron microscopy
PCR
Protein Assay
affinity

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