437 research outputs found
The VgrG Proteins Are "Ă la Carte" Delivery Systems for Bacterial Type VI Effectors
The bacterial type VI secretion system (T6SS) is a supra-molecular complex akin to bacteriophage tails, with VgrG proteins acting as a puncturing device. The Pseudomonas aeruginosa H1-T6SS has been extensively characterized. It is involved in bacterial killing and in the delivery of three toxins, Tse1â3. Here, we demonstrate the independent contribution of the three H1-T6SS co-regulated vgrG genes, vgrG1abc, to bacterial killing. A putative toxin is encoded in the vicinity of each vgrG gene, supporting the concept of specific VgrG/toxin couples. In this respect, VgrG1c is involved in the delivery of an Rhs protein, RhsP1. The RhsP1 C terminus carries a toxic activity, from which the producing bacterium is protected by a cognate immunity. Similarly, VgrG1a-dependent toxicity is associated with the PA0093 gene encoding a two-domain protein with a putative toxin domain (Toxin_61) at the C terminus. Finally, VgrG1b-dependent killing is detectable upon complementation of a triple vgrG1abc mutant. The VgrG1b-dependent killing is mediated by PA0099, which presents the characteristics of the superfamily nuclease 2 toxin members. Overall, these data develop the concept that VgrGs are indispensable components for the specific delivery of effectors. Several additional vgrG genes are encoded on the P. aeruginosa genome and are not linked genetically to other T6SS genes. A closer inspection of these clusters reveals that they also encode putative toxins. Overall, these associations further support the notion of an original form of secretion system, in which VgrG acts as the carrier
The rise of the Type VI secretion system
Bacterial cells have developed multiple strategies to communicate with their surrounding environment. The intracellular compartment is separated from the milieu by a relatively impermeable cell envelope through which small molecules can passively diffuse, while larger macromolecules, such as proteins, can be actively transported. In Gram-negative bacteria, the cell envelope is a double membrane, which houses several supramolecular protein complexes that facilitate the trafficking of molecules. For example, bacterial pathogens use these types of machines to deliver toxins into target eukaryotic host cells, thus subverting host cellular functions. Six different types of nanomachines, called Type I - Type VI secretion systems (T1SS - T6SS), can be readily identified by their composition and mode of action. A remarkable feature of these protein secretion systems is their similarity to systems with other biological functions, such as motility or the exchange of genetic material. The T6SS has provided a refreshing view on this concept since it shares similarity with the puncturing device of bacteriophages, which is used by these viruses to inject their DNA into bacterial target cells. In contrast, the bacterial T6SS transports toxins into other bacteria, engaging a ferocious competition for the colonization of their environment. Moreover, as with few other secretion systems, the T6SS is capable of injecting toxins into eukaryotic cells, which contributes to a successful infection. This highlights the multifunctional aspects of the T6SS, and our understanding of its mechanistic details is an intense field of investigation with significant implications for ecology, agriculture and medicine
Agrobacterium tumefaciens Deploys a Superfamily of Type VI Secretion DNase Effectors as Weapons for Interbacterial Competition In Planta
The type VI secretion system (T6SS) is a widespread molecular weapon deployed by many Proteobacteria to target effectors/toxins into both eukaryotic and prokaryotic cells. We report that Agrobacterium tumefaciens, a soil bacterium that triggers tumorigenesis in plants, produces a family of type VI DNase effectors (Tde) that are distinct from previously known polymorphic toxins and nucleases. Tde exhibits an antibacterial DNase activity that relies on a conserved HxxD motif and can be counteracted by a cognate immunity protein, Tdi. In vitro, A. tumefaciens T6SS could kill Escherichia coli but triggered a lethal counterattack by Pseudomonas aeruginosa upon injection of the Tde toxins. However, in an in planta coinfection assay, A. tumefaciens used Tde effectors to attack both siblings cells and P. aeruginosa to ultimately gain a competitive advantage. Such acquired T6SS-dependent fitness in vivo and conservation of Tde-Tdi couples in bacteria highlights a widespread antibacterial weapon beneficial for niche colonization
Biofilms and cyclic di-GMP (c-di-GMP) signaling: lessons from Pseudomonas aeruginosa and other bacteria
The cyclic di-GMP (c-di-GMP) second messenger represents a signaling system that regulates many bacterial behaviors and is of key importance for driving the lifestyle switch between motile loner cells and biofilm formers. This review provides an up-to-date compendium of c-di-GMP pathways connected to biofilm formation, biofilm-associated motilities, and other functionalities in the ubiquitous and opportunistic human pathogen Pseudomonas aeruginosa. This bacterium is frequently adopted as a model organism to study bacterial biofilm formation. Importantly, its versatility and adaptation capabilities are linked with a broad range of complex regulatory networks, including a large set of genes involved in c-di-GMP biosynthesis, degradation, and transmission
Evolution of Supermassive Black Holes from Cosmological Simulations
The correlations between the mass of supermassive black holes and properties
of their host galaxies are investigated through cosmological simulations. Black
holes grow from seeds of 100 solar masses inserted into density peaks present
in the redshift range 12-15. Seeds grow essentially by accreting matter from a
nuclear disk and also by coalescences resulting from merger episodes. At z=0,
our simulations reproduce the black hole mass function and the correlations of
the black hole mass both with stellar velocity dispersion and host dark halo
mass. Moreover, the evolution of the black hole mass density derived from the
present simulations agrees with that derived from the bolometric luminosity
function of quasars, indicating that the average accretion history of seeds is
adequately reproduced . However, our simulations are unable to form black holes
with masses above at , whose existence is inferred
from the bright quasars detected by the Sloan survey in this redshift range.Comment: Talk given at the International Workshop on Astronomy and
Relativistic Astrophysics (IWARA 2009), Maresias, Brazil. to be published in
the International Journal of Modern Physics
Coalescence Rate of Supermassive Black Hole Binaries Derived from Cosmological Simulations: Detection Rates for LISA and ET
The coalescence history of massive black holes has been derived from
cosmological simulations, in which the evolution of those objects and that of
the host galaxies are followed in a consistent way. The present study indicates
that supermassive black holes having masses greater than underwent up to 500 merger events along their history. The derived
coalescence rate per comoving volume and per mass interval permitted to obtain
an estimate of the expected detection rate distribution of gravitational wave
signals ("ring-down") along frequencies accessible by the planned
interferometers either in space (LISA) or in the ground (Einstein). For LISA,
in its original configuration, a total detection rate of about is
predicted for events having a signal-to-noise ratio equal to 10, expected to
occur mainly in the frequency range . For the Einstein gravitational
wave telescope, one event each 14 months down to one event each 4 years is
expected with a signal-to-noise ratio of 5, occurring mainly in the frequency
interval . The detection of these gravitational signals and their
distribution in frequency would be in the future an important tool able to
discriminate among different scenarios explaining the origin of supermassive
black holes.Comment: 18 pages, 7 figures, to appear in the IJMP
TagF-mediated repression of bacterial type VI secretion systems involves a direct interaction with the cytoplasmic protein Fha
The bacterial type VI secretion system (T6SS) delivers effectors into eukaryotic host cells or toxins into bacterial competitor for survival and fitness. The T6SS is positively regulated by the threonine phosphorylation pathway (TPP) and negatively by the T6SS-accessory protein TagF. Here, we studied the mechanisms underlying TagF-mediated T6SS repression in two distinct bacterial pathogens, Agrobacterium tumefaciens and Pseudomonas aeruginosa. We found that in A. tumefaciens, T6SS toxin secretion and T6SS-dependent antibacterial activity are suppressed by a two-domain chimeric protein consisting of TagF and PppA, a putative phosphatase. Remarkably, this TagF domain is sufficient to post-translationally repress the T6SS, and this inhibition is independent of TPP. This repression requires interaction with a cytoplasmic protein, Fha, critical for activating T6SS assembly. In P. aeruginosa, PppA and TagF are two distinct proteins that repress T6SS in a TPP-dependent and -independent pathways, respectively. P. aeruginosa TagF interacts with Fha1, suggesting that formation of this complex represents a conserved TagF-mediated regulatory mechanism. Using TagF variants with substitutions of conserved amino acid residues at predicted protein-protein interaction interfaces, we uncovered evidence that the TagF-Fha interaction is critical for TagF-mediated T6SS repression in both bacteria. TagF inhibits T6SS without affecting T6SS protein abundance in A. tumefaciens, but TagF overexpression reduces the protein levels of all analyzed T6SS components in P. aeruginosa. Our results indicate that TagF interacts with Fha, which in turn could impact different stages of T6SS assembly in different bacteria, possibly reflecting an evolutionary divergence in T6SS control
Manipulating the type VI secretion system spike to shuttle passenger proteins
The type VI secretion system (T6SS) is a contractile injection apparatus that translocates a spike loaded with various effectors directly into eukaryotic or prokaryotic target cells. Pseudomonas aeruginosa can load either one of its three T6SSs with a variety of toxic bullets using different but specific modes. The T6SS spike, which punctures the bacterial cell envelope allowing effector transport, consists of a torch-like VgrG trimer on which sits a PAAR protein sharpening the VgrG tip. VgrG itself sits on the Hcp tube and all elements, packed into a T6SS sheath, are propelled out of the cell and into target cells. On occasion, effectors are covalent extensions of VgrG, PAAR or Hcp proteins, which are then coined âevolvedâ components as opposed to canonical. Here, we show how various passenger domains could be fused to the C terminus of a canonical VgrG, VgrG1a from P. aeruginosa, and be sent into the bacterial culture supernatant. There is no restriction on the passenger type, although the efficacy may vary greatly, since we used either an unrelated T6SS protein, ÎČ-lactamase, a covalent extension of an âevolvedâ VgrG, VgrG2b, or a Hcp-dependent T6SS toxin, Tse2. Our data further highlights an exceptional modularity/flexibility for loading the T6SS nano-weapon. Refining the parameters to optimize delivery of passenger proteins of interest would have attractive medical and industrial applications. This may for example involve engineering the T6SS as a delivery system to shuttle toxins into either bacterial pathogens or tumour cells which would be an original approach in the fight against antimicrobial resistant bacteria or cancer
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