scholarly journals RhlR, but not RhlI, allowsP. aeruginosabacteria to evadeDrosophilaTep4-mediated opsonization

2017 ◽  
Author(s):  
Samantha Haller ◽  
Adrien Franchet ◽  
Abdul Hakkim ◽  
Jing Chen ◽  
Eliana Drenkard ◽  
...  
Keyword(s):  
Intestinal Barrier ◽  
Infection Model ◽  
Dependent Manner ◽  
Intestinal Infection ◽  
Wild Type ◽  
Independent Function ◽  
C Elegans ◽  
Injury Model ◽  
Mutant Phenotypes ◽  
Cellular Immune

ABSTRACTWhenDrosophilaflies feed onPseudomonas aeruginosastrain PA14, some bacteria cross the intestinal barrier and start proliferating inside the hemocoel. This process is limited by hemocytes through phagocytosis. We have previously shown that the PA14 quorum-sensing regulator RhlR is required for these bacteria to elude the cellular immune response. RhlI synthesizes the auto-inducer signal that activates RhlR. Here, we compare the null mutant phenotypes ofrhlRandrhlIin a variety of infection assays inDrosophilaand in the nematodeCaenorhabditis elegans. Surprisingly, inDrosophila, unlikeΔrhlRmutants,ΔrhlImutants are only modestly attenuated for virulence and are poorly phagocytosed and opsonized in a Thioester-containing Protein4-dependent manner. Likewise, ΔrhlIbut not ΔrhlRmutants colonize the digestive tract ofC. elegansand kill it as efficiently as wild-type PA14. Thus, RhlR has an RhlI-independent function in eluding detection or counter-acting the action of the immune system. In contrast to the intestinal infection model,Tep4mutant flies are more resistant to PA14 in a septic injury model, which also depends onrhlR. Thus, the Tep4 putative opsonin can either be protective or detrimental to host defense depending on the infection route.

scholarly journals The fliR gene contributes to the virulence of S. marcescens in a Drosophila intestinal infection model

2021 ◽  
Author(s):  
Bechara Sina Rahme ◽  
Matthieu Lestradet ◽  
Gisela Di Venanzio ◽  
Arshad Ayyaz ◽  
Miriam Wennida Yamba ◽  
...  
Keyword(s):  
Intestinal Epithelium ◽  
Intestinal Barrier ◽  
Body Cavity ◽  
Bacterial Virulence ◽  
The Body ◽  
Infection Model ◽  
Intestinal Infection ◽  
Intestinal Infections ◽  
Injury Model ◽  
Wide Range

Abstract Serratia marcescens is an opportunistic bacterium that infects a wide range of hosts including humans. It is a potent pathogen in a septic injury model of Drosophila melanogaster as five bacteria directly injected in the body cavity of the fly kill the host within a day. In contrast, flies do not succumb to ingested bacteria for days even though some bacteria traverse the intestinal barrier into the hemolymph within a couple of hours. The mechanisms by which S. marcescens attacks enterocytes and damages the intestinal epithelium remain uncharacterized. To better understand intestinal infections, we performed a genetic screen for loss of virulence of ingested S. marcescens in which we identified FliR, a structural component of the flagellum, as a virulence factor. Next, we compared the virulence of two flagellum mutants fliR and flhD using two Serratia strains. Both genes are required for S. marcescens to escape the gut lumen into the hemocoel indicating that the flagellum plays an important role for the passage of bacteria through the intestinal barrier. In contrast, fliR but not flhD is needed to severely damage the intestinal epithelium and ultimately kill the host. Our results therefore suggest a flagellum-independent role for fliR in bacterial virulence.

scholarly journals Goα Regulates Volatile Anesthetic Action in Caenorhabditis elegans

Genetics ◽  
2001 ◽  
Vol 158 (2) ◽  
pp. 643-655 ◽  
Author(s):  
Bruno van Swinderen ◽  
Laura B Metz ◽  
Laynie D Shebester ◽  
Jane E Mendel ◽  
Paul W Sternberg ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Novel Mutation ◽  
Volatile Anesthetic ◽  
Loss Of Function ◽  
Wild Type ◽  
Α Subunit ◽  
C Elegans ◽  
Missense Mutant ◽  
Anesthetic Action ◽  
Mutant Phenotypes

Abstract To identify genes controlling volatile anesthetic (VA) action, we have screened through existing Caenorhabditis elegans mutants and found that strains with a reduction in Go signaling are VA resistant. Loss-of-function mutants of the gene goa-1, which codes for the α-subunit of Go, have EC50s for the VA isoflurane of 1.7- to 2.4-fold that of wild type. Strains overexpressing egl-10, which codes for an RGS protein negatively regulating goa-1, are also isoflurane resistant. However, sensitivity to halothane, a structurally distinct VA, is differentially affected by Go pathway mutants. The RGS overexpressing strains, a goa-1 missense mutant found to carry a novel mutation near the GTP-binding domain, and eat-16(rf) mutants, which suppress goa-1(gf) mutations, are all halothane resistant; goa-1(null) mutants have wild-type sensitivities. Double mutant strains carrying mutations in both goa-1 and unc-64, which codes for a neuronal syntaxin previously found to regulate VA sensitivity, show that the syntaxin mutant phenotypes depend in part on goa-1 expression. Pharmacological assays using the cholinesterase inhibitor aldicarb suggest that VAs and GOA-1 similarly downregulate cholinergic neurotransmitter release in C. elegans. Thus, the mechanism of action of VAs in C. elegans is regulated by Goα, and presynaptic Goα-effectors are candidate VA molecular targets.

scholarly journals The Quorum-Sensing Molecule Autoinducer 2 Regulates Motility and Flagellar Morphogenesis in Helicobacter pylori

2007 ◽  
Vol 189 (17) ◽  
pp. 6109-6117 ◽  
Author(s):  
Bethany A. Rader ◽  
Shawn R. Campagna ◽  
Martin F. Semmelhack ◽  
Bonnie L. Bassler ◽  
Karen Guillemin
Keyword(s):  
Helicobacter Pylori ◽  
Quorum Sensing ◽  
Sigma Factor ◽  
Critical Role ◽  
Soft Agar ◽  
Dependent Manner ◽  
Flagellar Gene ◽  
Wild Type ◽  
Autoinducer 2 ◽  
Mutant Phenotypes

ABSTRACT The genome of the gastric pathogen Helicobacter pylori contains a homologue of the gene luxS, which has been shown to be responsible for production of the quorum-sensing signal autoinducer 2 (AI-2). We report here that deletion of the luxS gene in strain G27 resulted in decreased motility on soft agar plates, a defect that was complemented by a wild-type copy of the luxS gene and by the addition of cell-free supernatant containing AI-2. The flagella of the luxS mutant appeared normal; however, in genetic backgrounds lacking any of three flagellar regulators—the two-component sensor kinase flgS, the sigma factor σ28 (also called fliA), and the anti-sigma factor flgM—loss of luxS altered flagellar morphology. In all cases, the double mutant phenotypes were restored to the luxS + phenotype by the addition of synthetic 4,5-dihydroxy-2,3-pentanedione (DPD), which cyclizes to form AI-2. Furthermore, in all mutant backgrounds loss of luxS caused a decrease in transcript levels of the flagellar regulator flhA. Addition of DPD to luxS cells induced flhA transcription in a dose-dependent manner. Deletion of flhA in a wild-type or luxS mutant background resulted in identical loss of motility, flagella, and flagellar gene expression. These data demonstrate that AI-2 functions as a secreted signaling molecule upstream of FlhA and plays a critical role in global regulation of flagellar gene transcription in H. pylori.

scholarly journals Bacillus thuringiensis Spores and Vegetative Bacteria: Infection Capacity and Role of the Virulence Regulon PlcR Following Intrahaemocoel Injection of Galleria mellonella

Insects ◽  
2019 ◽  
Vol 10 (5) ◽  
pp. 129 ◽  
Author(s):  
Christophe Buisson ◽  
Michel Gohar ◽  
Eugénie Huillet ◽  
Christina Nielsen-LeRoux
Keyword(s):  
Bacillus Thuringiensis ◽  
Galleria Mellonella ◽  
Lethal Dose ◽  
Intestinal Barrier ◽  
Oral Route ◽  
Infection Model ◽  
Wild Type ◽  
Response Curves ◽  
Vegetative Cells ◽  
Crystal Toxins

Bacillus thuringiensis is an invertebrate pathogen that produces insecticidal crystal toxins acting on the intestinal barrier. In the Galleria mellonella larvae infection model, toxins from the PlcR virulence regulon contribute to pathogenicity by the oral route. While B. thuringiensis is principally an oral pathogen, bacteria may also reach the insect haemocoel following injury of the cuticle. Here, we address the question of spore virulence as compared to vegetative cells when the wild-type Bt407cry- strain and its isogenic ∆plcR mutant are inoculated directly into G. mellonella haemocoel. Mortality dose-response curves were constructed at 25 and 37 °C using spores or vegetative cell inocula, and the 50% lethal dose (LD50) in all infection conditions was determined after 48 h of infection. Our findings show that (i) the LD50 is lower for spores than for vegetative cells for both strains, while the temperature has no significant influence, and (ii) the ∆plcR mutant is four to six times less virulent than the wild-type strain in all infection conditions. Our results suggest that the environmental resistant spores are the most infecting form in haemocoel and that the PlcR virulence regulon plays an important role in toxicity when reaching the haemocoel from the cuticle and not only following ingestion.

scholarly journals A Chemokine-Degrading Extracellular Protease Made by Group A Streptococcus Alters Pathogenesis by Enhancing Evasion of the Innate Immune Response

2008 ◽  
Vol 76 (3) ◽  
pp. 978-985 ◽  
Author(s):  
Paul Sumby ◽  
Shizhen Zhang ◽  
Adeline R. Whitney ◽  
Fabiana Falugi ◽  
Guido Grandi ◽  
...  
Keyword(s):  
Immune Response ◽  
Innate Immune Response ◽  
Group A Streptococcus ◽  
Cell Envelope ◽  
Innate Immune ◽  
Infection Model ◽  
Dependent Manner ◽  
Wild Type ◽  
Group A

ABSTRACT Circumvention of the host innate immune response is critical for bacterial pathogens to infect and cause disease. Here we demonstrate that the group A Streptococcus (GAS; Streptococcus pyogenes) protease SpyCEP (S. pyogenes cell envelope protease) cleaves granulocyte chemotactic protein 2 (GCP-2) and growth-related oncogene alpha (GROα), two potent chemokines made abundantly in human tonsils. Cleavage of GCP-2 and GROα by SpyCEP abrogated their abilities to prime neutrophils for activation, detrimentally altering the innate immune response. SpyCEP expression is negatively regulated by the signal transduction system CovR/S. Purified recombinant CovR bound the spyCEP gene promoter region in vitro, indicating direct regulation. Immunoreactive SpyCEP protein was present in the culture supernatants of covR/S mutant GAS strains but not in supernatants from wild-type strains. However, wild-type GAS strains do express SpyCEP, where it is localized to the cell wall. Strain MGAS2221, an organism representative of the highly virulent and globally disseminated M1T1 GAS clone, differed significantly from its isogenic spyCEP mutant derivative strain in a mouse soft tissue infection model. Interestingly, and in contrast to previous studies, the isogenic mutant strain generated lesions of larger size than those formed following infection with the parent strain. The data indicate that SpyCEP contributes to GAS virulence in a strain- and disease-dependent manner.

scholarly journals The shelterin protein POT-1 anchors Caenorhabditis elegans telomeres through SUN-1 at the nuclear periphery

2013 ◽  
Vol 203 (5) ◽  
pp. 727-735 ◽  
Author(s):  
Helder C. Ferreira ◽  
Benjamin D. Towbin ◽  
Thibaud Jegou ◽  
Susan M. Gasser
Keyword(s):  
Caenorhabditis Elegans ◽  
Nuclear Periphery ◽  
Telomere Maintenance ◽  
Dependent Manner ◽  
Wild Type ◽  
Dna Structures ◽  
C Elegans ◽  
Differentiated Cells ◽  
Sumo Ligase ◽  
Early Embryos

Telomeres are specialized protein–DNA structures that protect chromosome ends. In budding yeast, telomeres form clusters at the nuclear periphery. By imaging telomeres in embryos of the metazoan Caenorhabditis elegans, we found that telomeres clustered only in strains that had activated an alternative telomere maintenance pathway (ALT). Moreover, as in yeast, the unclustered telomeres in wild-type embryos were located near the nuclear envelope (NE). This bias for perinuclear localization increased during embryogenesis and persisted in differentiated cells. Telomere position in early embryos required the NE protein SUN-1, the single-strand binding protein POT-1, and the small ubiquitin-like modifier (SUMO) ligase GEI-17. However, in postmitotic larval cells, none of these factors individually were required for telomere anchoring, which suggests that additional mechanisms anchor in late development. Importantly, targeted POT-1 was sufficient to anchor chromatin to the NE in a SUN-1–dependent manner, arguing that its effect at telomeres is direct. This high-resolution description of telomere position within C. elegans extends our understanding of telomere organization in eukaryotes.

scholarly journals Cryptococcus neoformans Gene Involved in Mammalian Pathogenesis Identified by a Caenorhabditis elegans Progeny-Based Approach

2005 ◽  
Vol 73 (12) ◽  
pp. 8219-8225 ◽  
Author(s):  
Robin J. Tang ◽  
Julia Breger ◽  
Alexander Idnurm ◽  
Kimberly J. Gerik ◽  
Jennifer K. Lodge ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Caenorhabditis Elegans ◽  
Cryptococcus Neoformans ◽  
Fungal Pathogen ◽  
Infection Model ◽  
Microbial Pathogenesis ◽  
Wild Type ◽  
C Elegans ◽  
Growth Defects ◽  
High Throughput Method

ABSTRACT Caenorhabditis elegans can serve as a substitute host for the study of microbial pathogenesis. We found that mutations in genes of the fungal pathogen Cryptococcus neoformans involved in mammalian virulence allow C. elegans to produce greater numbers of progeny than when exposed to wild-type fungus. We used this property to screen a library of C. neoformans mutants for strains that permit larger C. elegans brood sizes. In this screen, we identified a gene homologous to Saccharomyces cerevisiae ROM2. C. neoformans rom2 mutation resulted in a defect in mating and growth defects at elevated temperature or in the presence of cell wall or hyperosmolar stresses. An effect of the C. neoformans rom2 mutation in virulence was confirmed in a murine inhalation infection model. We propose that a screen for progeny-permissive mutants of microorganisms can serve as a high-throughput method for identifying novel loci related to mammalian pathogenesis.

scholarly journals Regulation of body length and male tail ray pattern formation of Caenorhabditis elegans by a member of TGF-beta family

Development ◽  
1999 ◽  
Vol 126 (6) ◽  
pp. 1337-1347 ◽  
Author(s):  
K. Morita ◽  
K.L. Chow ◽  
N. Ueno
Keyword(s):  
Caenorhabditis Elegans ◽  
Body Length ◽  
Target Genes ◽  
Dependent Manner ◽  
Tgf Beta ◽  
Wild Type ◽  
Male Tail ◽  
C Elegans ◽  
Sensory Ray ◽  
Epistasis Analysis

We have identified a new member of the TGF-beta superfamily, CET-1, from Caenorhabditis elegans, which is expressed in the ventral nerve cord and other neurons. cet-1 null mutants have shortened bodies and male tail abnormal phenotype resembling sma mutants, suggesting cet-1, sma-2, sma-3 and sma-4 share a common pathway. Overexpression experiments demonstrated that cet-1 function requires wild-type sma genes. Interestingly, CET-1 appears to affect body length in a dose-dependent manner. Heterozygotes for cet-1 displayed body lengths ranging between null mutant and wild type, and overexpression of CET-1 in wild-type worms elongated body length close to lon mutants. In male sensory ray patterning, lack of cet-1 function results in ray fusions. Epistasis analysis revealed that mab-21 lies downstream and is negatively regulated by the cet-1/sma pathway in the male tail. Our results show that cet-1 controls diverse biological processes during C. elegans development probably through different target genes.

scholarly journals CaMK (CMK-1) and O-GlcNAc transferase (OGT-1) modulate mechanosensory responding and habituation in an interstimulus interval-dependent manner in Caenorhabditis elegans

2017 ◽  
Author(s):  
Evan L. Ardiel ◽  
Troy A. McDiarmid ◽  
Tiffany A. Timbers ◽  
Kirsten C. Y. Lee ◽  
Javad Safaei ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Interstimulus Interval ◽  
Behavioral Plasticity ◽  
Neural Circuit ◽  
Model Organisms ◽  
Dependent Manner ◽  
Mechanical Stimuli ◽  
C Elegans ◽  
Mutant Phenotypes ◽  
Glcnac Transferase

AbstractThe ability to learn is an evolutionarily conserved adaptation that remains incompletely understood. Genetically tractable model organisms facilitate mechanistic explanations of learning that span genetic, neural circuit, and behavioural levels. Many aspects of neural physiology, including processes that underlie learning (e.g. neurotransmitter release and long-lasting changes in synaptic strength), are regulated by brief and local changes in [μm] levels of free intracellular Ca2+. On this scale, changes in [Ca2+] activate many Ca2+-sensors, including the Ca2+/calmodulin-dependent kinases (CaMKs). Here we reveal that the Caenorhabditis elegans ortholog of CaMK1/4, CMK-1, functions in primary sensory neurons to regulate responses to mechanical stimuli and behavioral plasticity, specifically habituation, a conserved form of non-associative learning. The habituation phenotypes of cmk-1 mutants were dependent on interstimulus interval (ISI), such that CMK-1 slows habituation at short ISIs, but promotes it at long ISIs. We predicted potential CaMK phosphorylation targets from catalytic site analysis of the human and C. elegans CaMKs and mutant analysis of these candidates implicated O-linked N-acetylglucosamine (O-GlcNAc) transferase, OGT-1, in mechanosensitivity and learning. Cell specific rescue and knockdown experiments showed that both CMK-1 and OGT-1 function cell autonomously in mechanosensory neurons to modulate learning. Interestingly, despite their similar mutant phenotypes, detailed behavioral analysis of double mutants demonstrated that CMK-1 and OGT-1 act in parallel genetic pathways. Our research identifies CMK-1 and OGT-1 as co-expressed yet independent regulators of mechanosensitivity and learning.

scholarly journals HSP90 buffers newly induced mutations in massively mutated plant lines

2018 ◽  
Author(s):  
G. Alex Mason ◽  
Keisha D Carlson ◽  
Maximilian O Press ◽  
Kerry L Bubb ◽  
Christine Queitsch
Keyword(s):  
Genetic Variation ◽  
De Novo ◽  
Embryonic Lethality ◽  
Model Organisms ◽  
Dependent Manner ◽  
Wild Type ◽  
Induced Mutations ◽  
Emergent Feature ◽  
Phenotypic Robustness ◽  
Mutant Phenotypes

AbstractRobustness to both genetic and environmental change is an emergent feature of living systems. Loss of phenotypic robustness can be associated with increased penetrance of genetic variation. In model organisms and in humans, the phenotypic consequences of standing genetic variation can be buffered by the molecular chaperone HSP90. However, it has been argued that HSP90 has the opposite effect on newly introduced genetic variation. To test the buffering effect of HSP90 on new mutations, we introduced vast numbers of mutations into wild-type and HSP90-reduced plants and assessed embryonic lethality and early seedling phenotypes for thousands of offspring. Although the levels of newly introduced mutations were similar in the two backgrounds, the HSP90-reduced plants showed a significantly greater frequency of embryonic lethality and severe phenotypic abnormalities, consistent with higher penetrance and expressivity of newly introduced genetic variation. We further demonstrate that some mutant phenotypes were heritable in an HSP90-dependent manner, and we map candidate HSP90-dependent polymorphisms. Moreover, both sequence and phenotypic analyses of wild-type and HSP90-reduced plants suggest that the HSP90-dependent phenotypes are largely due the newly introduced mutations rather than to an increased mutation rate in HSP90-reduced plants. Taken together, our results support a model in which HSP90 buffers newly introduced mutations, and the phenotypic consequences of such mutations outweigh those of mutations arising de novo in response to HSP90 perturbation.

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