Brucella Modulates Secretory Trafficking via Multiple Type IV Secretion Effector Proteins

Sebenzile Myeni, Robert Child, Tony W. Ng, John J. Kupko, Tara D. Wehrly, Stephen F. Porcella, Leigh A. Knodler, Jean Celli

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

The intracellular pathogenic bacterium Brucella generates a replicative vacuole (rBCV) derived from the endoplasmic reticulum via subversion of the host cell secretory pathway. rBCV biogenesis requires the expression of the Type IV secretion system (T4SS) VirB, which is thought to translocate effector proteins that modulate membrane trafficking along the endocytic and secretory pathways. To date, only a few T4SS substrates have been identified, whose molecular functions remain unknown. Here, we used an in silico screen to identify putative T4SS effector candidate proteins using criteria such as limited homology in other bacterial genera, the presence of features similar to known VirB T4SS effectors, GC content and presence of eukaryotic-like motifs. Using β-lactamase and CyaA adenylate cyclase reporter assays, we identified eleven proteins translocated into host cells by Brucella, five in a VirB T4SS-dependent manner, namely BAB1_0678 (BspA), BAB1_0712 (BspB), BAB1_0847 (BspC), BAB1_1671 (BspE) and BAB1_1948 (BspF). A subset of the translocated proteins targeted secretory pathway compartments when ectopically expressed in HeLa cells, and the VirB effectors BspA, BspB and BspF inhibited protein secretion. Brucella infection also impaired host protein secretion in a process requiring BspA, BspB and BspF. Single or combined deletions of bspA, bspB and bspF affected Brucella ability to replicate in macrophages and persist in the liver of infected mice. Taken together, these findings demonstrate that Brucella modulates secretory trafficking via multiple T4SS effector proteins that likely act coordinately to promote Brucella pathogenesis.

Original languageEnglish (US)
Article numbere1003556
JournalPLoS Pathogens
Volume9
Issue number8
DOIs
StatePublished - Aug 2013

Fingerprint

Brucella
Secretory Pathway
Proteins
Base Composition
Vacuoles
HeLa Cells
Adenylyl Cyclases
Endoplasmic Reticulum
Computer Simulation
Membrane Proteins
Macrophages
Bacteria
Liver
Infection

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology
  • Medicine(all)

Cite this

Myeni, S., Child, R., Ng, T. W., Kupko, J. J., Wehrly, T. D., Porcella, S. F., ... Celli, J. (2013). Brucella Modulates Secretory Trafficking via Multiple Type IV Secretion Effector Proteins. PLoS Pathogens, 9(8), [e1003556]. https://doi.org/10.1371/journal.ppat.1003556

Brucella Modulates Secretory Trafficking via Multiple Type IV Secretion Effector Proteins. / Myeni, Sebenzile; Child, Robert; Ng, Tony W.; Kupko, John J.; Wehrly, Tara D.; Porcella, Stephen F.; Knodler, Leigh A.; Celli, Jean.

In: PLoS Pathogens, Vol. 9, No. 8, e1003556, 08.2013.

Research output: Contribution to journalArticle

Myeni, S, Child, R, Ng, TW, Kupko, JJ, Wehrly, TD, Porcella, SF, Knodler, LA & Celli, J 2013, 'Brucella Modulates Secretory Trafficking via Multiple Type IV Secretion Effector Proteins', PLoS Pathogens, vol. 9, no. 8, e1003556. https://doi.org/10.1371/journal.ppat.1003556
Myeni, Sebenzile ; Child, Robert ; Ng, Tony W. ; Kupko, John J. ; Wehrly, Tara D. ; Porcella, Stephen F. ; Knodler, Leigh A. ; Celli, Jean. / Brucella Modulates Secretory Trafficking via Multiple Type IV Secretion Effector Proteins. In: PLoS Pathogens. 2013 ; Vol. 9, No. 8.
@article{45934a6f7f6b4208aee42f2ce3fb9fbd,
title = "Brucella Modulates Secretory Trafficking via Multiple Type IV Secretion Effector Proteins",
abstract = "The intracellular pathogenic bacterium Brucella generates a replicative vacuole (rBCV) derived from the endoplasmic reticulum via subversion of the host cell secretory pathway. rBCV biogenesis requires the expression of the Type IV secretion system (T4SS) VirB, which is thought to translocate effector proteins that modulate membrane trafficking along the endocytic and secretory pathways. To date, only a few T4SS substrates have been identified, whose molecular functions remain unknown. Here, we used an in silico screen to identify putative T4SS effector candidate proteins using criteria such as limited homology in other bacterial genera, the presence of features similar to known VirB T4SS effectors, GC content and presence of eukaryotic-like motifs. Using β-lactamase and CyaA adenylate cyclase reporter assays, we identified eleven proteins translocated into host cells by Brucella, five in a VirB T4SS-dependent manner, namely BAB1_0678 (BspA), BAB1_0712 (BspB), BAB1_0847 (BspC), BAB1_1671 (BspE) and BAB1_1948 (BspF). A subset of the translocated proteins targeted secretory pathway compartments when ectopically expressed in HeLa cells, and the VirB effectors BspA, BspB and BspF inhibited protein secretion. Brucella infection also impaired host protein secretion in a process requiring BspA, BspB and BspF. Single or combined deletions of bspA, bspB and bspF affected Brucella ability to replicate in macrophages and persist in the liver of infected mice. Taken together, these findings demonstrate that Brucella modulates secretory trafficking via multiple T4SS effector proteins that likely act coordinately to promote Brucella pathogenesis.",
author = "Sebenzile Myeni and Robert Child and Ng, {Tony W.} and Kupko, {John J.} and Wehrly, {Tara D.} and Porcella, {Stephen F.} and Knodler, {Leigh A.} and Jean Celli",
year = "2013",
month = "8",
doi = "10.1371/journal.ppat.1003556",
language = "English (US)",
volume = "9",
journal = "PLoS Pathogens",
issn = "1553-7366",
publisher = "Public Library of Science",
number = "8",

}

TY - JOUR

T1 - Brucella Modulates Secretory Trafficking via Multiple Type IV Secretion Effector Proteins

AU - Myeni, Sebenzile

AU - Child, Robert

AU - Ng, Tony W.

AU - Kupko, John J.

AU - Wehrly, Tara D.

AU - Porcella, Stephen F.

AU - Knodler, Leigh A.

AU - Celli, Jean

PY - 2013/8

Y1 - 2013/8

N2 - The intracellular pathogenic bacterium Brucella generates a replicative vacuole (rBCV) derived from the endoplasmic reticulum via subversion of the host cell secretory pathway. rBCV biogenesis requires the expression of the Type IV secretion system (T4SS) VirB, which is thought to translocate effector proteins that modulate membrane trafficking along the endocytic and secretory pathways. To date, only a few T4SS substrates have been identified, whose molecular functions remain unknown. Here, we used an in silico screen to identify putative T4SS effector candidate proteins using criteria such as limited homology in other bacterial genera, the presence of features similar to known VirB T4SS effectors, GC content and presence of eukaryotic-like motifs. Using β-lactamase and CyaA adenylate cyclase reporter assays, we identified eleven proteins translocated into host cells by Brucella, five in a VirB T4SS-dependent manner, namely BAB1_0678 (BspA), BAB1_0712 (BspB), BAB1_0847 (BspC), BAB1_1671 (BspE) and BAB1_1948 (BspF). A subset of the translocated proteins targeted secretory pathway compartments when ectopically expressed in HeLa cells, and the VirB effectors BspA, BspB and BspF inhibited protein secretion. Brucella infection also impaired host protein secretion in a process requiring BspA, BspB and BspF. Single or combined deletions of bspA, bspB and bspF affected Brucella ability to replicate in macrophages and persist in the liver of infected mice. Taken together, these findings demonstrate that Brucella modulates secretory trafficking via multiple T4SS effector proteins that likely act coordinately to promote Brucella pathogenesis.

AB - The intracellular pathogenic bacterium Brucella generates a replicative vacuole (rBCV) derived from the endoplasmic reticulum via subversion of the host cell secretory pathway. rBCV biogenesis requires the expression of the Type IV secretion system (T4SS) VirB, which is thought to translocate effector proteins that modulate membrane trafficking along the endocytic and secretory pathways. To date, only a few T4SS substrates have been identified, whose molecular functions remain unknown. Here, we used an in silico screen to identify putative T4SS effector candidate proteins using criteria such as limited homology in other bacterial genera, the presence of features similar to known VirB T4SS effectors, GC content and presence of eukaryotic-like motifs. Using β-lactamase and CyaA adenylate cyclase reporter assays, we identified eleven proteins translocated into host cells by Brucella, five in a VirB T4SS-dependent manner, namely BAB1_0678 (BspA), BAB1_0712 (BspB), BAB1_0847 (BspC), BAB1_1671 (BspE) and BAB1_1948 (BspF). A subset of the translocated proteins targeted secretory pathway compartments when ectopically expressed in HeLa cells, and the VirB effectors BspA, BspB and BspF inhibited protein secretion. Brucella infection also impaired host protein secretion in a process requiring BspA, BspB and BspF. Single or combined deletions of bspA, bspB and bspF affected Brucella ability to replicate in macrophages and persist in the liver of infected mice. Taken together, these findings demonstrate that Brucella modulates secretory trafficking via multiple T4SS effector proteins that likely act coordinately to promote Brucella pathogenesis.

UR - http://www.scopus.com/inward/record.url?scp=84883317837&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84883317837&partnerID=8YFLogxK

U2 - 10.1371/journal.ppat.1003556

DO - 10.1371/journal.ppat.1003556

M3 - Article

C2 - 23950720

AN - SCOPUS:84883317837

VL - 9

JO - PLoS Pathogens

JF - PLoS Pathogens

SN - 1553-7366

IS - 8

M1 - e1003556

ER -