YscP, a Yersinia protein required for Yop secretion that is surface exposed, and released in low Ca2+

I Stainier, S Bleves, C Josenhans, L Karmani, C Kerbourch, I Lambermont, S Tötemeyer, A Boyd, G R Cornelis

Research output: Contribution to journalArticle

Abstract

The Yersinia Ysc apparatus is made of more than 20 proteins, 11 of which have homologues in many type III systems. Here, we characterize YscP from Yersinia enterocolitica. This 515-residue protein has a high proline content, a large tandem repetition and a slow migration in SDS-PAGE. Unlike the products of neighbouring genes, it has a counterpart only in Pseudomonas aeruginosa and it varies even between Yersinia Ysc machineries. An yscPDelta97-465 mutant was unable to secrete any Yop, even under conditions overcoming feedback inhibition of Yop synthesis. Interestingly, a cloned yscPDelta57-324 from Yersinia pestis introduced in the yscPDelta97-465 mutant can sustain a significant Yop secretion and thus partially complemented the mutation. This explains the leaky phenotype observed with the yscP mutant of Y. pestis. In accordance with this secretion deficiency, YscP is required for the delivery of Yop effectors into macrophages. Mechanical shearing, immunolabelling and electron microscopy experiments showed that YscP is exposed at the bacterial surface when bacteria are incubated at 37 degrees C in the presence of Ca2+ and thus do not secrete Yops. At 37 degrees C, when Ca2+ ions are chelated, YscP is released like a Yop protein. We conclude that YscP is a part of the Ysc injectisome which is localized at the bacterial surface and is destabilized by Ca2+ chelation.
Original languageEnglish
Pages (from-to)1005-18
Number of pages14
JournalMolecular Microbiology
Volume37
Issue number5
Publication statusPublished - 2000

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Yersinia
Yersinia pestis
Yersinia enterocolitica
Proteins
Proline
Pseudomonas aeruginosa
Polyacrylamide Gel Electrophoresis
Electron Microscopy
Macrophages
Ions
Bacteria
Phenotype
Mutation
Genes

Cite this

Stainier, I., Bleves, S., Josenhans, C., Karmani, L., Kerbourch, C., Lambermont, I., ... Cornelis, G. R. (2000). YscP, a Yersinia protein required for Yop secretion that is surface exposed, and released in low Ca2+. Molecular Microbiology, 37(5), 1005-18.
Stainier, I ; Bleves, S ; Josenhans, C ; Karmani, L ; Kerbourch, C ; Lambermont, I ; Tötemeyer, S ; Boyd, A ; Cornelis, G R. / YscP, a Yersinia protein required for Yop secretion that is surface exposed, and released in low Ca2+. In: Molecular Microbiology. 2000 ; Vol. 37, No. 5. pp. 1005-18.
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Stainier, I, Bleves, S, Josenhans, C, Karmani, L, Kerbourch, C, Lambermont, I, Tötemeyer, S, Boyd, A & Cornelis, GR 2000, 'YscP, a Yersinia protein required for Yop secretion that is surface exposed, and released in low Ca2+', Molecular Microbiology, vol. 37, no. 5, pp. 1005-18.

YscP, a Yersinia protein required for Yop secretion that is surface exposed, and released in low Ca2+. / Stainier, I; Bleves, S; Josenhans, C; Karmani, L; Kerbourch, C; Lambermont, I; Tötemeyer, S; Boyd, A; Cornelis, G R.

In: Molecular Microbiology, Vol. 37, No. 5, 2000, p. 1005-18.

Research output: Contribution to journalArticle

TY - JOUR

T1 - YscP, a Yersinia protein required for Yop secretion that is surface exposed, and released in low Ca2+

AU - Stainier, I

AU - Bleves, S

AU - Josenhans, C

AU - Karmani, L

AU - Kerbourch, C

AU - Lambermont, I

AU - Tötemeyer, S

AU - Boyd, A

AU - Cornelis, G R

PY - 2000

Y1 - 2000

N2 - The Yersinia Ysc apparatus is made of more than 20 proteins, 11 of which have homologues in many type III systems. Here, we characterize YscP from Yersinia enterocolitica. This 515-residue protein has a high proline content, a large tandem repetition and a slow migration in SDS-PAGE. Unlike the products of neighbouring genes, it has a counterpart only in Pseudomonas aeruginosa and it varies even between Yersinia Ysc machineries. An yscPDelta97-465 mutant was unable to secrete any Yop, even under conditions overcoming feedback inhibition of Yop synthesis. Interestingly, a cloned yscPDelta57-324 from Yersinia pestis introduced in the yscPDelta97-465 mutant can sustain a significant Yop secretion and thus partially complemented the mutation. This explains the leaky phenotype observed with the yscP mutant of Y. pestis. In accordance with this secretion deficiency, YscP is required for the delivery of Yop effectors into macrophages. Mechanical shearing, immunolabelling and electron microscopy experiments showed that YscP is exposed at the bacterial surface when bacteria are incubated at 37 degrees C in the presence of Ca2+ and thus do not secrete Yops. At 37 degrees C, when Ca2+ ions are chelated, YscP is released like a Yop protein. We conclude that YscP is a part of the Ysc injectisome which is localized at the bacterial surface and is destabilized by Ca2+ chelation.

AB - The Yersinia Ysc apparatus is made of more than 20 proteins, 11 of which have homologues in many type III systems. Here, we characterize YscP from Yersinia enterocolitica. This 515-residue protein has a high proline content, a large tandem repetition and a slow migration in SDS-PAGE. Unlike the products of neighbouring genes, it has a counterpart only in Pseudomonas aeruginosa and it varies even between Yersinia Ysc machineries. An yscPDelta97-465 mutant was unable to secrete any Yop, even under conditions overcoming feedback inhibition of Yop synthesis. Interestingly, a cloned yscPDelta57-324 from Yersinia pestis introduced in the yscPDelta97-465 mutant can sustain a significant Yop secretion and thus partially complemented the mutation. This explains the leaky phenotype observed with the yscP mutant of Y. pestis. In accordance with this secretion deficiency, YscP is required for the delivery of Yop effectors into macrophages. Mechanical shearing, immunolabelling and electron microscopy experiments showed that YscP is exposed at the bacterial surface when bacteria are incubated at 37 degrees C in the presence of Ca2+ and thus do not secrete Yops. At 37 degrees C, when Ca2+ ions are chelated, YscP is released like a Yop protein. We conclude that YscP is a part of the Ysc injectisome which is localized at the bacterial surface and is destabilized by Ca2+ chelation.

M3 - Article

C2 - 10972820

VL - 37

SP - 1005

EP - 1018

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 5

ER -

Stainier I, Bleves S, Josenhans C, Karmani L, Kerbourch C, Lambermont I et al. YscP, a Yersinia protein required for Yop secretion that is surface exposed, and released in low Ca2+. Molecular Microbiology. 2000;37(5):1005-18.