VgrG-5 is a Burkholderia type VI secretion system-exported protein required for multinucleated giant cell formation and virulence.
The type VI secretion system (T6SS) has emerged as a critical virulence factor for the group of closely related Burkholderia spp. that includes Burkholderia pseudomallei, B. mallei, and B. thailandensis. While the genomes of these bacteria, referred to as the Bptm group, appear to encode several T6SSs, we and others have shown that one of these, type VI secretion system 5 (T6SS-5), is required for virulence in mammalian infection models. Despite its pivotal role in the pathogenesis of the Bptm group, the effector repertoire of T6SS-5 has remained elusive. Here we used quantitative mass spectrometry to compare the secretome of wild-type B. thailandensis to that of a mutant harboring a nonfunctional T6SS-5. This analysis identified VgrG-5 as a novel secreted protein whose export depends on T6SS-5 function. Bioinformatics analysis revealed that VgrG-5 is a specialized VgrG protein that harbors a C-terminal domain (CTD) conserved among Bptm group species. We found that a vgrG-5 ΔCTD mutant is avirulent in mice and is unable to stimulate the fusion of host cells, a hallmark of the Bptm group previously shown to require T6SS-5 function. The singularity of VgrG-5 as a detected T6SS-5 substrate, taken together with the essentiality of its CTD for virulence, suggests that the protein is critical for the effector activity of T6SS-5. Intriguingly, we show that unlike the bacterial-cell-targeting T6SSs characterized so far, T6SS-5 localizes to the bacterial cell pole. We propose a model whereby the CTD of VgrG-5-, propelled by T6SS-5-, plays a key role in inducing membrane fusion, either by the recruitment of other factors or by direct participation.
Department of Microbiology, University of Washington, Seattle, Washington, USA.
Sandra
S
Schwarz
Pragya
P
Singh
Johanna D
JD
Robertson
Michele
M
LeRoux
Shawn J
SJ
Skerrett
David R
DR
Goodlett
T Eoin
TE
West
Joseph D
JD
Mougous
D000818:Animals; D058947:Bacterial Secretion Systems / Q000502:physiology*; D015153:Blotting, Western; D019117:Burkholderia / Q000378:metabolism / Q000472:pathogenicity*; D002478:Cells, Cultured; D015726:Giant Cells / Q000378:metabolism / Q000502:physiology*; D006790:Host-Parasite Interactions / Q000502:physiology; D008264:Macrophages / Q000378:metabolism; D013058:Mass Spectrometry; D008561:Membrane Fusion / Q000502:physiology; D051379:Mice; D008856:Microscopy, Fluorescence; D014774:Virulence / Q000235:genetics / Q000502:physiology; D037521:Virulence Factors / Q000378:metabolism
D058947:Bacterial Secretion Systems; D037521:Virulence Factors