Function and molecular architecture of the Yersinia injectisome tip complex

Petr Broz; Catherine A Mueller; Shirley A Müller; Ansgar Philippsen; Isabel Sorg; Andreas Engel; Guy R Cornelis

doi:10.1111/j.1365-2958.2007.05871.x

Function and molecular architecture of the Yersinia injectisome tip complex

Petr Broz, Catherine A Mueller, Shirley A Müller, Ansgar Philippsen, Isabel Sorg, Andreas Engel, Guy R Cornelis

Research output: Contribution to journal › Article › peer-review

Abstract

By quantitative immunoblot analyses and scanning transmission electron microscopy (STEM), we determined that the needle of the Yersinia enterocolitica E40 injectisome consists of 139 +/- 19 YscF subunits and that the tip complex is formed by three to five LcrV monomers. A pentamer represented the best fit for an atomic model of this complex. The N-terminal globular domain of LcrV forms the base of the tip complex, while the central globular domain forms the head. Hybrids between LcrV and its orthologues PcrV (Pseudomonas aeruginosa) or AcrV (Aeromonas salmonicida) were engineered and recombinant Y. enterocolitica expressing the different hybrids were tested for their capacity to form the translocation pore by a haemolysis assay. There was a good correlation between haemolysis, insertion of YopB into erythrocyte membranes and interaction between YopB and the N-terminal globular domain of the tip complex subunit. Hence, the base of the tip complex appears to be critical for the functional insertion of YopB into the host cell membrane.

Original language	English
Pages (from-to)	1311-20
Number of pages	10
Journal	Molecular Microbiology
Volume	65
Issue number	5
DOIs	https://doi.org/10.1111/j.1365-2958.2007.05871.x
Publication status	Published - 2007

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title = "Function and molecular architecture of the Yersinia injectisome tip complex",

abstract = "By quantitative immunoblot analyses and scanning transmission electron microscopy (STEM), we determined that the needle of the Yersinia enterocolitica E40 injectisome consists of 139 +/- 19 YscF subunits and that the tip complex is formed by three to five LcrV monomers. A pentamer represented the best fit for an atomic model of this complex. The N-terminal globular domain of LcrV forms the base of the tip complex, while the central globular domain forms the head. Hybrids between LcrV and its orthologues PcrV (Pseudomonas aeruginosa) or AcrV (Aeromonas salmonicida) were engineered and recombinant Y. enterocolitica expressing the different hybrids were tested for their capacity to form the translocation pore by a haemolysis assay. There was a good correlation between haemolysis, insertion of YopB into erythrocyte membranes and interaction between YopB and the N-terminal globular domain of the tip complex subunit. Hence, the base of the tip complex appears to be critical for the functional insertion of YopB into the host cell membrane.",

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T1 - Function and molecular architecture of the Yersinia injectisome tip complex

AU - Broz, Petr

AU - Mueller, Catherine A

AU - Müller, Shirley A

AU - Philippsen, Ansgar

AU - Sorg, Isabel

AU - Engel, Andreas

AU - Cornelis, Guy R

PY - 2007

Y1 - 2007

N2 - By quantitative immunoblot analyses and scanning transmission electron microscopy (STEM), we determined that the needle of the Yersinia enterocolitica E40 injectisome consists of 139 +/- 19 YscF subunits and that the tip complex is formed by three to five LcrV monomers. A pentamer represented the best fit for an atomic model of this complex. The N-terminal globular domain of LcrV forms the base of the tip complex, while the central globular domain forms the head. Hybrids between LcrV and its orthologues PcrV (Pseudomonas aeruginosa) or AcrV (Aeromonas salmonicida) were engineered and recombinant Y. enterocolitica expressing the different hybrids were tested for their capacity to form the translocation pore by a haemolysis assay. There was a good correlation between haemolysis, insertion of YopB into erythrocyte membranes and interaction between YopB and the N-terminal globular domain of the tip complex subunit. Hence, the base of the tip complex appears to be critical for the functional insertion of YopB into the host cell membrane.

AB - By quantitative immunoblot analyses and scanning transmission electron microscopy (STEM), we determined that the needle of the Yersinia enterocolitica E40 injectisome consists of 139 +/- 19 YscF subunits and that the tip complex is formed by three to five LcrV monomers. A pentamer represented the best fit for an atomic model of this complex. The N-terminal globular domain of LcrV forms the base of the tip complex, while the central globular domain forms the head. Hybrids between LcrV and its orthologues PcrV (Pseudomonas aeruginosa) or AcrV (Aeromonas salmonicida) were engineered and recombinant Y. enterocolitica expressing the different hybrids were tested for their capacity to form the translocation pore by a haemolysis assay. There was a good correlation between haemolysis, insertion of YopB into erythrocyte membranes and interaction between YopB and the N-terminal globular domain of the tip complex subunit. Hence, the base of the tip complex appears to be critical for the functional insertion of YopB into the host cell membrane.

U2 - 10.1111/j.1365-2958.2007.05871.x

DO - 10.1111/j.1365-2958.2007.05871.x

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SN - 0950-382X

VL - 65

SP - 1311

EP - 1320

JO - Molecular Microbiology

JF - Molecular Microbiology

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ER -

Function and molecular architecture of the Yersinia injectisome tip complex

Abstract

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