In Vivo Transcriptional Profiling of Listeria monocytogenes and Mutagenesis Identify New Virulence Factors Involved in Infection

Listeria monocytogenes is a human intracellular pathogen able to colonize host tissues after ingestion of contaminated food, causing severe invasive infections. In order to gain a better understanding of the nature of host–pathogen interactions, we studied the L. monocytogenes genome expression during mouse infection. In the spleen of infected mice, ≈20% of the Listeria genome is differentially expressed, essentially through gene activation, as compared to exponential growth in rich broth medium. Data presented here show that, during infection, Listeria is in an active multiplication phase, as revealed by the high expression of genes involved in replication, cell division and multiplication. In vivo bacterial growth requires increased expression of genes involved in adaptation of the bacterial metabolism and stress responses, in particular to oxidative stress. Listeria interaction with its host induces cell wall metabolism and surface expression of virulence factors. During infection, L. monocytogenes also activates subversion mechanisms of host defenses, including resistance to cationic peptides, peptidoglycan modifications and release of muramyl peptides. We show that the in vivo differential expression of the Listeria genome is coordinated by a complex regulatory network, with a central role for the PrfA-SigB interplay. In particular, L. monocytogenes up regulates in vivo the two major virulence regulators, PrfA and VirR, and their downstream effectors. Mutagenesis of in vivo induced genes allowed the identification of novel L. monocytogenes virulence factors, including an LPXTG surface protein, suggesting a role for S-layer glycoproteins and for cadmium efflux system in Listeria virulence

Stabilin-1 plays a protective role against Listeria monocytogenes infection through the regulation of cytokine and chemokine production and immune cell recruitment

Scavenger receptors are part of a complex surveillance system expressed by host cells to efficiently orchestrate innate immune response against bacterial infections. Stabilin-1 (STAB-1) is a scavenger receptor involved in cell trafficking, inflammation, and cancer; however, its role in infection remains to be elucidated. Listeria monocytogenes (Lm) is a major intracellular human food-borne pathogen causing severe infections in susceptible hosts. Using a mouse model of infection, we demonstrate here that STAB-1 controls Lm-induced cytokine and chemokine production and immune cell accumulation in Lm-infected organs. We show that STAB-1 also regulates the recruitment of myeloid cells in response to Lm infection and contributes to clear circulating bacteria. In addition, whereas STAB-1 appears to promote bacterial uptake by macrophages, infection by pathogenic Listeria induces the down regulation of STAB-1 expression and its delocalization from the host cell membrane.We propose STAB-1 as a new SR involved in the control of Lm infection through the regulation of host defense mechanisms, a process that would be targeted by bacterial virulence factors to promote infection

A bacterial protein targets the BAHD1 chromatin complex to stimulate type III interferon response

International audienceIntracellular pathogens such as $Listeria\ monocytogenes$ subvert cellular functions through the interaction of bacterial effectors with host components. Here we found that a secreted listerial virulence factor, LntA, could target the chromatin repressor BAHD1 in the host cell nucleus to activate interferon IFN-stimulated genes (ISGs). IFN-$\lambda$ expression was induced in response to infection of epithelial cells with bacteria lacking LntA; however, the BAHD1-chromatin associated complex repressed downstream ISGs. In contrast, in cells infected with $lntA$-expressing bacteria, LntA prevented BAHD1 recruitment to ISGs and stimulated their expression. Murine listeriosis decreased in BAHD1$^{+/-}$ mice or when $lntA$ was constitutively expressed. Thus, the LntA-BAHD1 interplay may modulate IFN-$\lambda$-mediated immune response to control bacterial colonization of the host

The specific surface area and chemical composition of diamond dust near Barrow, Alaska

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/95687/1/jgrd17349.pd

MouR controls the expression of the Listeria monocytogenes Agr system and mediates virulence

The foodborne pathogen Listeria monocytogenes (Lm) causes invasive infection in susceptible ani- mals and humans. To survive and proliferate within hosts, this facultative intracellular pathogen tightly coordinates the expression of a complex regulatory network that controls the expression of virulence fac- tors. Here, we identified and characterized MouR, a novel virulence regulator of Lm. Through RNA-seq transcriptomic analysis, we determined the MouR regulon and demonstrated how MouR positively con- trols the expression of the Agr quorum sensing sys- tem (agrBDCA) of Lm. The MouR three-dimensional structure revealed a dimeric DNA-binding transcrip- tion factor belonging to the VanR class of the GntR superfamily of regulatory proteins. We also showed that by directly binding to the agr promoter region, MouR ultimately modulates chitinase activity and biofilm formation. Importantly, we demonstrated by in vitro cell invasion assays and in vivo mice infec- tions the role of MouR in Lm virulence.Peer reviewe

IDENTIFICATION ET CARACTERISATION DE GENES DE SINORHIZOBIUM MELILOTI REGULES AU COURS DE L'INTERACTION SYMBIOTIQUE AVEC MEDICAGO SATIVA

SINORHIZOBIUM MELILOTI INDUIT SUR LES RACINES DE LA LUZERNE LA FORMATION D'ORGANES SPECIALISES DANS LA FIXATION DE L'AZOTE, LES NODOSITES. LE DEVELOPPEMENT DE CES NODOSITES NECESSITE L'EXPRESSION COORDONNEE DE NOMBREUX GENES VEGETAUX ET BACTERIENS. AFIN D'IDENTIFIER DE TELS GENES, POTENTIELLEMENT IMPORTANTS POUR L'ETABLISSEMENT DE LA SYMBIOSE, NOUS NOUS SOMMES INTERESSE AUX GENES BACTERIENS DONT L'EXPRESSION EST REGULEE DANS LES NODOSITES. NOUS AVONS ISOLE PAR RNA FINGERPRINTING BY ARBITRARILY PRIMED PCR 17 ADNC DIFFERENTIELLEMENT EXPRIMES EN CONDITIONS SYMBIOTIQUES PAR RAPPORT AUX CONDITIONS DE CULTURE PURE. 13 ADNC CORRESPONDENT A DES GENES REGULES POSITIVEMENT EN SYMBIOSE ALORS QUE 4 SONT REPRIMES EN CONDITIONS SYMBIOTIQUES. L'ANALYSE DETAILLEE DE L'EXPRESSION DES GENES CORRESPONDANTS NOUS A PERMIS DE METTRE EN EVIDENCE UNE VARIETE INSOUPCONNEE DE PROFILS DE REGULATION. 7 ADNC CORRESPONDENT A DES GENES NIF ET FIX CONNUS OU PREDITS. LES PRODUITS DE 4 ADNC PRESENTENT UNE FORTE SIMILARITE AVEC DES PROTEINES NON IDENTIFIEES CHEZ S. MELILOTI, A SAVOIR UN COMPOSANT DU COMPLEXE PYRUVATE DESHYDROGENASE, UNE PROTEINE DE SURFACE DE LA CELLULE, UN TRANSPORTEUR DE CUIVRE ET UNE ARGININOSUCCINATE LYASE. LES PRODUITS DES 6 AUTRES ADNC NE MONTRENT AUCUNE SIMILARITE AVEC DES PROTEINES CONNUES. UN DES ADNC CODE POUR UN DES COMPOSES DU COMPLEXE PYRUVATE DESHYDROGENASE, UN ELEMENT CLE DU METABOLISME CARBONE QUI CATALYSE LA FORMATION D'ACETYL-COA A PARTIR DE PYRUVATE. NOUS AVONS MONTRE QUE L'EXPRESSION DE PDHA EST TRES FORTEMENT INDUITE EN SYMBIOSE ET EST INDEPENDANTE DE LA CASCADE QUI REGULE LES GENES DE FIXATION DE L'AZOTE ET LES GENES DE RESPIRATION DE S. MELILOTI. L'EXPRESSION DE PDHA EST INDUITE EN CULTURE PURE PAR LE PYRUVATE. CETTE INDUCTION AINSI QUE L'ACTIVATION SYMBIOTIQUE DES GENES PDH ONT LIEU AU MEME PROMOTEUR. CES RESULTATS SUGGERENT UN ROLE ESSENTIEL DE LA PYRUVATE DESHYDROGENASE POUR LE METABOLISME BACTERIEN SYMBIOTIQUE ET UN ROLE REGULATEUR DU PYRUVATE EN SYMBIOSE.TOULOUSE3-BU Sciences (315552104) / SudocSudocFranceF

Auto, a surface associated autolysin of Listeria monocytogenes required for entry into eukaryotic cells and virulence

International audienceListeria monocytogenes is an opportunistic food-borne human and animal pathogen. Several surface proteins expressed by this intracellular pathogen are critical for the infectious process. By in silico analysis we compared the surface protein repertories of L. monocytogenes and of the non-pathogenic species Listeria innocua and identified a gene encoding a surface protein of L. monocytogenes absent in L. innocua. This gene that we named aut encodes a protein (Auto) of 572 amino acids containing a signal sequence, a N-terminal autolysin domain and a C-terminal cell wall-anchoring domain made up of four GW modules. We show here that the aut gene is expressed independently of the virulence gene regulator PrfA and encodes a surface protein with an autolytic activity. We provide evidence that Auto is required for entry of L. monocytogenes into cultured non-phagocytic eukaryotic cells. The low invasiveness of an aut deletion mutant correlates with its reduced virulence following intravenous inoculation of mice and oral infection of guinea pigs. During infection, the autolytic activity of Auto may also be critical. Auto appears thus as a novel type of L. monocytogenes virulence factor

Control of cytoskeletal dynamics during cellular responses to pore forming toxins

Following damage by pore forming toxins (PFTs) host cells engage repair processes and display profound cytoskeletal remodeling and concomitant plasma membrane (PM) blebbing. We have recently demonstrated that host cells utilize similar mechanisms to control cytoskeletal dynamics in response to PFTs and during cell migration. This involves assembly of cortical actomyosin bundles, reorganisation of the endoplasmic reticulum (ER) network, and the interaction between the ER chaperone Gp96 and the molecular motor Non-muscle Myosin Heavy Chain IIA (NMHCIIA). Consequently, Gp96 regulates actomyosin activity, PM blebbing and cell migration, and protects PM integrity against PFTs. In addition, we observed that PFTs increase association of Gp96 and ER vacuoles with the cell surface or within PM blebs loosely attached to the cell body. Similarly, gut epithelial cells damaged by PFTs in vivo were shown to release microvilli structures or directly purge cytoplasmic content. Cytoplasmic purging involves profound cytoskeletal remodeling and ER vacuolation, suggesting that our observations recapitulate recovery processes in vivo. Here, we discuss our findings in light of the current understanding of PM repair mechanisms and in vivo recovery responses to PFTs

Mechanisms protecting host cells against bacterial pore-forming toxins

Pore-forming toxins (PFTs) are key virulence determinants produced and secreted by a variety of human bacterial pathogens. They disrupt the plasma membrane (PM) by generating stable protein pores, which allow uncontrolled exchanges between the extracellular and intracellular milieus, dramatically disturbing cellular homeostasis. In recent years, many advances were made regarding the characterization of conserved repair mechanisms that allow eukaryotic cells to recover from mechanical disruption of the PM membrane. However, the specificities of the cell recovery pathways that protect host cells against PFT-induced damage remain remarkably elusive. During bacterial infections, the coordinated action of such cell recovery processes defines the outcome of infected cells and is, thus, critical for our understanding of bacterial pathogenesis. Here, we review the cellular pathways reported to be involved in the response to bacterial PFTs and discuss their impact in single-cell recovery and infection