The potassium impermeable apical membrane of insect epithelia: a target for development of safe pesticides

Harvey, William R.; Crawford, Dwight N.; Eisen, N.; Fernandes, Veronica F.; Spaeth, Daniel D.; Wolfersberger, Michael G.

doi:10.1590/s0074-02761987000700008

Cited by 6 publications

(1 citation statement)

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“…Insect midguts were dissected from silkworm larvae and used to prepare the total proteins of midgut brush border membrane vesicles (BBMVs) by differential precipitation using MgCl 2 [ 62 ] and stored at -80°C until use.…”

Section: Methodsmentioning

confidence: 99%

Bacillus bombysepticus α-Toxin Binding to G Protein-Coupled Receptor Kinase 2 Regulates cAMP/PKA Signaling Pathway to Induce Host Death

Lin

Cheng

et al. 2016

PLoS Pathog

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Bacterial pathogens and their toxins target host receptors, leading to aberrant behavior or host death by changing signaling events through subversion of host intracellular cAMP level. This is an efficient and widespread mechanism of microbial pathogenesis. Previous studies describe toxins that increase cAMP in host cells, resulting in death through G protein-coupled receptor (GPCR) signaling pathways by influencing adenylyl cyclase or G protein activity. G protein-coupled receptor kinase 2 (GRK2) has a central role in regulation of GPCR desensitization. However, little information is available about the pathogenic mechanisms of toxins associated with GRK2. Here, we reported a new bacterial toxin-Bacillus bombysepticus (Bb) α-toxin that was lethal to host. We showed that Bb α-toxin interacted with BmGRK2. The data demonstrated that Bb α-toxin directly bound to BmGRK2 to promote death by affecting GPCR signaling pathways. This mechanism involved stimulation of Gαs, increase level of cAMP and activation of protein kinase A (PKA). Activated cAMP/PKA signal transduction altered downstream effectors that affected homeostasis and fundamental biological processes, disturbing the structural and functional integrity of cells, resulting in death. Preventing cAMP/PKA signaling transduction by inhibitions (NF449 or H-89) substantially reduced the pathogenicity of Bb α-toxin. The discovery of a toxin-induced host death specifically linked to GRK2 mediated signaling pathway suggested a new model for bacterial toxin action. Characterization of host genes whose expression and function are regulated by Bb α-toxin and GRK2 will offer a deeper understanding of the pathogenesis of infectious diseases caused by pathogens that elevate cAMP.

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Section: Methodsmentioning

confidence: 99%

Bacillus bombysepticus α-Toxin Binding to G Protein-Coupled Receptor Kinase 2 Regulates cAMP/PKA Signaling Pathway to Induce Host Death

Lin

Cheng

et al. 2016

PLoS Pathog

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The crystal δ‐endotoxins of Bacillus thuringiensis: Models for their mechanism of action on the insect gut

1993

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The crystal 6-endotoxins of Bacillus thuringiensis (Bt) are a family of insecticidal proteins which have been known for some time to kill insects by lysing their gut epithelial cells, but the precise molecular mechanism of toxicity has remained elusive. The recent publication of the crystal structure of a Bt &endotoxin has made it possible for us to model the molecular events that occur as the toxin binds to its receptor and inserts into the membrane to form a pore. Using our knowledge of insect gut physiology, we can also predict the effect on the insect of the formation of a toxic pore. We present a new model to explain the events that occur in the insect gut during toxin action.

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Bacillus thuringiensis δ-Endotoxin Proteins Show a Correlation in Toxicity and Short Circuit Current Inhibition Against Helicoverpa zea

Karim

Dean

2000

Current Microbiology

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Pesticidal activity of Bacillus thuringiensis delta-endotoxins, Cry1Aa, Cry1Ab, Cry1Ac, and Cry2A, was determined by using the force-feeding bioassay method to 4(th) instar larvae of Helicoverpa zea. H. zea was susceptible to Bt toxins in the order Cry1Ac > Cry1Ab > Cry1Aa > Cry2A with 63.60, 89.04, 159.65, and 375.78 ng/larvae respectively. The abilities of selected Bacillus thuringiensis toxins to inhibit short circuit current (I(SC)) in midgut epithelia of H. zea were also investigated by voltage clamp assay. The voltage-clamp studies were conducted on isolated midguts, measuring the inhibition of short circuit current (I(SC)) by activated toxin. A Cry1Aa toxin dilution of 33.3 and 500 ng/ml resulted in inhibition of I(SC) of -2.29 microA/min (lag time 15 min) and -4.48 microA/min (lag time, 2 min) respectively. The Cry1Ab dilution of 25 ng/ml inhibited I(SC) to -1.39 microA/min, a lag time of 14 min, and 333.3 ng/ml dilution resulted in decay of I(SC) -2.49 microA/min, lag time 1 min respectively. The Cry1Ac lower dilution 16.7 ng/ml inhibited I(SC) to -1.39 microA/min, lag time 4 min, and a high dilution 333.3 ng/ml decay I(SC) to -2.44 microA/min, lag time 1 min. The inhibition of I(SC) (-1.10 microA/min, lag time 25) at lower dilution (33.3 ng/ml) and high dilution (500 ng/ml), decay (-2.38 microA/min, lag time 5 min), showed a correlation between toxin concentration and inhibitory response with Cry2A toxin. The lag time decreased with increasing concentration of toxin applied, which is additional evidence of dose response besides direct correlation of toxicity assays and I(SC).

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The potassium impermeable apical membrane of insect epithelia: a target for development of safe pesticides

Cited by 6 publications

References 0 publications

Bacillus bombysepticus α-Toxin Binding to G Protein-Coupled Receptor Kinase 2 Regulates cAMP/PKA Signaling Pathway to Induce Host Death

Bacillus bombysepticus α-Toxin Binding to G Protein-Coupled Receptor Kinase 2 Regulates cAMP/PKA Signaling Pathway to Induce Host Death

The crystal δ‐endotoxins of Bacillus thuringiensis: Models for their mechanism of action on the insect gut

Bacillus thuringiensis δ-Endotoxin Proteins Show a Correlation in Toxicity and Short Circuit Current Inhibition Against Helicoverpa zea

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