The type IV secretion system core component VirB8 interacts via the β1‐strand with VirB10

Sharifahmadian, Mahzad; Nlend, Ingrid Um; Lecoq, Lauriane; Omichinski, James G.; Baron, Christian

doi:10.1002/1873-3468.12770

Cited by 7 publications

(12 citation statements)

References 36 publications

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“…2 B ). These results are in accordance with our previous data, suggesting that the fusion proteins mutually stabilize each other and are degraded in the absence of an interaction in the heterologous host Escherichia coli (10, 20). …”

Section: Resultssupporting

confidence: 93%

“…We used phage display and the bacterial two-hybrid (BTH) assay to identify interaction sites between Brucella VirB proteins (10, 15, 19). The information on protein-protein interaction sites is then being applied to test the importance of the interactions between the homologs in T4SS for which more biological readouts are available such as A. tumefaciens and pKM101 (20). Following up on our previous work that identified the N terminus as interaction site (10), we determined which residues of VirB6 from B. suis (VirB6b) are involved in the interaction with VirB10b.…”

Section: Resultsmentioning

confidence: 99%

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Interaction via the N terminus of the type IV secretion system (T4SS) protein VirB6 with VirB10 is required for VirB2 and VirB5 incorporation into T-pili and for T4SS function

Mary

Fouillen

Bessette

et al. 2018

Journal of Biological Chemistry

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Many bacterial pathogens employ multicomponent protein complexes such as type IV secretion systems (T4SSs) to transfer virulence factors into host cells. Here we studied the interaction between two essential T4SS components: the very hydrophobic inner membrane protein VirB6, which may be a component of the translocation channel, and VirB10, which links the inner and outer bacterial membranes. To map the interaction site between these two T4SS components, we conducted alanine scanning and deleted six-amino acid stretches from the N-terminal periplasmic domain of VirB6 from Brucella suis. Using the bacterial two-hybrid system to analyze the effects of these alterations on the VirB6–VirB10 interaction, we identified the amino acid regions 16–21 and 28–33 and Leu-18 in VirB6 as being required for this interaction. SDS-PAGE coupled with Western blotting of cell lysates and native PAGE of detergent-extracted membrane proteins revealed that the corresponding VirB6 residues in Agrobacterium tumefaciens (Phe-20 and amino acids 18–23 and 30–35) modulate the stability of both VirB6 and VirB5. However, the results from immuno-EM and super-resolution microscopy suggested that these regions and residues are not required for membrane association or for polar localization of VirB6. The six-amino acid deletions in the N terminus of VirB6 abolished pilus formation and virulence of A. tumefaciens, and the corresponding deletions in the VirB6 homolog TraD from the plasmid pKM101-T4SS abrogated plasmid transfer. Our results indicate that specific residues of the VirB6 N-terminal domain are required for VirB6 stabilization, its interaction with VirB10, and the incorporation of VirB2 and VirB5 into T-pili.

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Section: Resultssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Interaction via the N terminus of the type IV secretion system (T4SS) protein VirB6 with VirB10 is required for VirB2 and VirB5 incorporation into T-pili and for T4SS function

Mary

Fouillen

Bessette

et al. 2018

Journal of Biological Chemistry

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“…These indicate a structure that makes contact with TrwG/VirB8 peri and that is flexibly (shown in dashed lines) connected to another structure that makes contact with the TrwI/VirB6 TM helices and with the two subunits of the TrwK/VirB4 dimer. The density was too poor to be assigned but could correspond to TrwE/ VirB10 NTD , which is known to not only make a major part of the OMCC but also has an IM TM and a cytoplasmic tail that, in other T4SSs, has been known to interact with VirB8, VirB6 and VirB4 [70][71][72] . However, it could be that this stretch of density may correspond to different proteins.…”

Section: Data Availabilitymentioning

confidence: 99%

Cryo-EM structure of a type IV secretion system

Macé

Vadakkepat

Redzej

et al. 2022

Nature

127

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Bacterial conjugation is the fundamental process of unidirectional transfer of DNAs, often plasmid DNAs, from a donor cell to a recipient cell1. It is the primary means by which antibiotic resistance genes spread among bacterial populations2,3. In Gram-negative bacteria, conjugation is mediated by a large transport apparatus—the conjugative type IV secretion system (T4SS)—produced by the donor cell and embedded in both its outer and inner membranes. The T4SS also elaborates a long extracellular filament—the conjugative pilus—that is essential for DNA transfer4,5. Here we present a high-resolution cryo-electron microscopy (cryo-EM) structure of a 2.8 megadalton T4SS complex composed of 92 polypeptides representing 8 of the 10 essential T4SS components involved in pilus biogenesis. We added the two remaining components to the structural model using co-evolution analysis of protein interfaces, to enable the reconstitution of the entire system including the pilus. This structure describes the exceptionally large protein–protein interaction network required to assemble the many components that constitute a T4SS and provides insights on the unique mechanism by which they elaborate pili.

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“…It has demonstrated that Omp19 is a possible vaccine candidate and has adjuvant activity as protease inhibitor, which can be used as positive control [ 49 , 50 ]. VirB8 from Brucella is a bitopic inner membrane protein, which undergoes several protein–protein interactions that have an impact on both the functionality and assembly of the T4SS complex, suggesting a key role in the T4S system (Figure 5 B) [ 51 , 52 ]. HlyD is a component of the prototypical alpha-hemolysin (HlyA) bacterial type I secretion T1S system, along with the other components HlyB and TolC (Figure 5 C) [ 53 ].…”

Section: Resultsmentioning

confidence: 99%

Screening of potential vaccine candidates against pathogenic Brucella spp. using compositive reverse vaccinology

Zai

Yin

Guo

et al. 2021

Vet Res

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Brucella spp. are Gram-negative, facultative intracellular bacteria that cause brucellosis in humans and various animals. The threat of brucellosis has increased, yet currently available live attenuated vaccines still have drawbacks. Therefore, subunit vaccines, produced using protein antigens and having the advantage of being safe, cost-effective and efficacious, are urgently needed. In this study, we used core proteome analysis and a compositive RV methodology to screen potential broad-spectrum antigens against 213 pathogenic strains of Brucella spp. with worldwide geographic distribution. Candidate proteins were scored according to six biological features: subcellular localization, antigen similarity, antigenicity, mature epitope density, virulence, and adhesion probability. In the RV analysis, a total 32 candidate antigens were picked out. Of these, three proteins were selected for assessment of immunogenicity and preliminary protection in a mouse model: outer membrane protein Omp19 (used as a positive control), type IV secretion system (T4SS) protein VirB8, and type I secretion system (T1SS) protein HlyD. These three antigens with a high degree of conservation could induce specific humoral and cellular immune responses. Omp19, VirB8 and HlyD could substantially reduce the organ bacterial load of B. abortus S19 in mice and provide varying degrees of protection. In this study, we demonstrated the effectiveness of this unique strategy for the screening of potential broad-spectrum antigens against Brucella. Further evaluation is needed to identify the levels of protection conferred by the vaccine antigens against wild-type pathogenic Brucella species challenge.

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The type IV secretion system core component VirB8 interacts via the β1‐strand with VirB10

Cited by 7 publications

References 36 publications

Interaction via the N terminus of the type IV secretion system (T4SS) protein VirB6 with VirB10 is required for VirB2 and VirB5 incorporation into T-pili and for T4SS function

Interaction via the N terminus of the type IV secretion system (T4SS) protein VirB6 with VirB10 is required for VirB2 and VirB5 incorporation into T-pili and for T4SS function

Cryo-EM structure of a type IV secretion system

Screening of potential vaccine candidates against pathogenic Brucella spp. using compositive reverse vaccinology

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