Activation of adenylate cyclase by forskolin increases the protein stability of RCAN1 (DSCR1 or Adapt78)

Overexpression of Regulator of Calcineurin 1 (RCAN1/DSCR1/Adapt78) is known to inhibit the calcineurin-NFAT dependent signaling pathway. In this report, we find that activation of adenylate cyclase by forskolin increases the expression of RCAN1 through the increase of the protein’s half-life. The ability of forskolin to increase the accumulation of RCAN1 protein is significantly inhibited with protein kinase A inhibitors such as KT5720 and H-89. Furthermore, forskolin targets the central and C-terminal region of RCAN1 and enhances the inhibitory effect of RCAN1 on the calcineurin-mediated activation of NFAT. Our findings provide the first evidence that the accumulation of the RCAN1 protein by cAMP acts as an important regulatory mechanism in the control of the calcineurin-dependent cellular pathway.

Structured summary

MINT-7262390: PKA (uniprotkb:P22694) phosphorylates (MI:0217) RCAN1 (uniprotkb:P53805) by protein kinase assay (MI:0424)     

Identification of EGF as an Important Regulator for Promoting CYP3A4 Expression in Human Embryonic Stem Cell-Derived Hepatocytes Using TALEN-Based Gene Targeting

Functional human hepatocytes are one of the most significant tools for studying drug absorption, distribution, metabolism and excretion/toxicity （ADME/Tox）, especially for applications in preclinical drug development （Sahi et al., 2010; Godoy et al., 2013）. They provide the closest in vitro model to the human liver and the only model that mimics the drug metabolic profiles found in vivo. However, these cells lose their metabolic function rapidly and dramatically during the in vitro culture process, which largely hinders their wider application in drug development （Sahi et al., 2010; Godoy et al., 2013）. To overcome this obstacle, it is important to regulate the activities of key genes which are responsible for the detoxification metabolic function of human hepatocytes.     

CFTR: a missing link between exocrine and endocrine pancreas?

<正>Cystic fibrosis(CF),a life-shortening hereditary disease mainly afflicting people of Caucasian origins,is caused by loss-of-function mutations in the CFTR(Cystic Fibrosis Transmembrane conductance Regulator)gene,which encodes a phosphorylation-activated,but ATP-gated anion channel expressed primarily in epithelial cells.To date,nearly 2000 mutations have been identified as pathogenic,     

Phenological models of development in Pandemis heparana and Adoxophyes orana for timing the application of Insect Growth Regulators with juvenile-hormone activity

Control of leafrollers using Insect Growth Regulators with juvenile-hormone acitivity (IGR) is a major issue in research on Integrated Pest Management in apple orchards. The IGR is applied at the time of emergence of the last-larval instar of leafrollers, thus causing a disturbance of metamorphosis.Simulation models on the development of Pandemis heparana (Denn. et Schiff.) and Adoxophyes orana (F.v.R.) were developed, partly on the basis of experiments, partly on data from literature, to predict the time of emergence of the various stages, particularly of the last-larval instar and the pupa. The models use the state-variable approach, and include only temperature as a driving variable.Simulated curves of emergence of last-instar larvae, pupae and moths corresponded well with observations on field-collected larvae, reared to adult stage in an outdoor insectary. The curves of pheromone trap catches showed a delay relative to the calculated and observed curves for the eclosion of pupae.To investigate whether the time of IGR application could be related to a temperature sum, the relation between emergence curves of last-instar larvae and temperature sums was studied for several years. For this purpose simulated curves were used, because observations on emergence of last-instar covered only a few years.

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Résumé Le contrôle des Tordeuses par les régulateurs de croissance d'Insectes (IGR), à activité d'hormone juvénile, est un progrès majeur dans la recherche pour la lutte intégrée contre les ennemis des vergers de pommiers.L'IGR appliqué lors de l'apparition du dernier stade larvaire des Tordeuses, perturbe ainsi la métamorphose ultérieure. Afin de prévoir le moment d'apparition des différents stades, et plus particulièrement du dernier stade larvaire, des modèles de simulation du développement de Pandemis heparana (Denn. et Schiff.) et Adoxophyes orana (F.v.R.) ont été élaborés à partir d'expériences et en se basant sur des résultats de la littérature.Le modèle est basé sur l'examen de la variable d'état, la température étant la variable discriminante suivie.Les courbes simulées d'apparition des larves de dernier stade, des nymphes et des papillons correspondent aux observations faites sur les larves récoltées dans la nature et élevées jusqu'au stade adulte dans un insectarium en plein air.Le taux de capture dans les pièges à phéromone présente un retard par rapport aux courbes calculées et observées pour l'éclosion des nymphes. Différentes causes peuvent être attribuées à cette différence, mais l'influence d'aucune d'elles n'a été étudiée.Afin de déterminer si le moment de l'application d'IGR peut être lié à une somme de températures, les courbes d'apparition des larves de dernier stade ont été étudiées en relation avec les sommes de températures portant sur plusieurs années. Les observations concernant l'apparition des larves de dernier stade ne couvrant que 2 années, des courbes simulées ont été utilisées à cet effet. L'emploid de la relation liant le moment de l'apparition à la somme des températures permet de n'avoir à calculer que les sommes de températures pour déterminer le moment opportun de l'application des IGR.

     

Extracts of Marine cyanobacteria stimulated somatic embryogenesis of Daucus carota L.

Twenty five strains of marine cyanobacteria were screened for their ability to promote carrot somatic embryogenesis. Hot water extracts prepared from 21 of these strains promoted plantlet formation. Extracts from four strains increased plantlet numbers to an average of over 3.7-fold. Dialysates and nondialysates of each of these extracts also increased plantlet formation. For extracts from filamentous cyanobacteria, Nostoc sp. and Anabaena sp., dialysate was more effective (4.2-fold increase) than nondialysate (3.0-fold increase), whereas for unicellular strains Synechococcus sp. and Xenococcus sp., nondialysate was more effective (5.2-fold increase) than the dialysate (3.2-fold increase). These cyanobacterial extracts also promoted embryolike structure formation from two-year old carrot cell cultures which were unable to produce plantlets using the usual methods. Here, we demonstrate the existence in marine cyanobacterial extracts of low and high molecular weight factors which strongly promote somatic embryogenesis in carrot cell cultures.Abbreviations MS Murashige and Skoog medium - 2, 4-D 2, 4-dichlorophenoxyacetic acid - PCV packed cell volume     

A Subset of Cytokinin Two-component Signaling System Plays a Role in Cold Temperature Stress Response in Arabidopsis

A multistep two-component signaling system is established as a key element of cytokinin signaling in Arabidopsis. Here, we provide evidence for a function of the two-component signaling system in cold stress response in Arabidopsis. Cold significantly induced the expression of a subset of A-type ARR genes and of GUS in Pro_ARR7:GUS transgenic Arabidopsis. AHK2 and AHK3 were found to be primarily involved in mediating cold to express A-type ARRs despite cytokinin deficiency. Cold neither significantly induced AHK2 and AHK3 expression nor altered the cytokinin contents of wild type within the 4 h during which the A-type ARR genes exhibited peak expression in response to cold, indicating that cold might induce ARR expression via the AHK2 and AHK3 proteins without alterations in cytokinin levels. The ahk2 ahk3 and ahk3 ahk4 mutants exhibited enhanced freezing tolerance compared with wild type. These ahk double mutants acclimated as efficiently to cold as did wild type. The overexpression of the cold-inducible ARR7 in Arabidopsis resulted in a hypersensitivity response to freezing temperatures under cold-acclimated conditions. The expression of C-repeat/dehydration-responsive element target genes was not affected by ARR7 overexpression as well as in ahk double mutants. By contrast, the arr7 mutants showed increased freezing tolerance. The ahk2 ahk3 and arr7 mutants showed hypersensitive response to abscisic acid (ABA) for germination, whereas ARR7 overexpression lines exhibited insensitive response to ABA. These results suggest that AHK2 and AHK3 and the cold-inducible A-type ARRs play a negative regulatory role in cold stress signaling via inhibition of ABA response, occurring independently of the cold acclimation pathway.     

Dexras1 inhibits adenylyl cyclase

Dexras1 is a steroid hormone-induced Ras family G protein that acts as a receptor-independent activator of signaling by Gi/o family heterotrimeric G proteins. We examined the effects of Dexras1 on the activity of adenylyl cylase, a target of inhibitory regulation by Gialpha x GTP. Constitutively active Gsalpha (Q227L) increased cAMP levels 43-fold above baseline, and Dexras1 expression inhibited cAMP levels by 61% (P < 0.01). Dexras1 mediated inhibition of adenylyl cyclase was blocked by treatment pertussis toxin or by co-expression of RGS4, but was not inhibited by with dominant-interfering (G203T or G204A) mutants of Gi alpha2. Dexras1 decreased forskolin-stimulated CREB activation (P < 0.01) and this activity was also inhibited by co-expression of RGS4. These findings indicate that Dexras1 expression leads to ligand-independent activation of both Gialpha- and G(beta)gamma-dependent arms of the Gi signaling cascade, and suggest that Dexras1 may exert physiologically relevant inhibitory effects on the cAMP-PKA-CREB.     

Effects of triflumuron on the two-spotted spider mite,Tetranychus urticae (Acari: Tetranichydae)

The effects of triflumuron on the mortality, fecundity, and fertility of the two-spotted spider mite, Tetranychus urticae, were evaluated in the laboratory. No differences in toxicity for larvae, protonymphs and deutonymphs were observed, but immature stages were 3.8-times more susceptible than adults at the LC₅₀. The compound exhibited a direct contact ovicidal activity influenced by eggs age. 48–72 hold eggs were significantly more sensitive than eggs of the other age classes. No hatch inhibition was observed in eggs laid by treated adult females using a sublethal dose at two physiological times (<12 and 48–72h old). However, fecundity decreased in younger treated females, but it increased in the older-ones. The toxicity for immatures and eggs, and the sublethal effects described suggest that triflumuron could be an interesting incorporation in integrated pest programs of T. urticae. This revised version was published online in July 2006 with corrections to the Cover Date.     

Mosquito Control Evaluations of an Insect Growth Regulator, Pyriproxyfen against Culex pipiens pattens (Diptera, Culicidae) Larvae in Marsh Area, Korea

ABSTRACT Mosquito control evaluations of the granular formulation of 0.5% pyriproxyfen were undertaken in marshes and a pond in Busan during the period of July through September, 2001. In marsh condition where Culex pipiens pattens were predominantly breeding, the treatment of pyriproxyfen produced over an average of 95% mosquito larval reduction at a concentration of 0.05 mg/1 for the first 4 weeks in spite of heavy precipitation (total 274.4 mm) during the period. Reduction rates of 72.6% and 32.3% appeared in the 5th and 7th weeks probably due to heavy input of sewage and flood. However, the reduction rate increased again at the 8th week, showing a 94.5% reduction, probably because of the slow releasing of pyriproxyfen granular formulation. In the second treatment of pyriproxyfen, complete mortality was obtained at the 1st and 3rd weeks. Also, a satisfactory level of Cx. pipiens larval reduction of 96.6% was observed at the 4th week. In a pond assessment, the treatment of pyriproxyfen at a concentration of 0.05 mg/1 produced a complete reduction of Cx. pipiens pattens from the 1st week through the 4th week after treatment. In the second treatment, 100.0% reduction was obtained at the 1st and 2nd weeks in the same sites. The treatment resulted in 84.4% and 97.9% reduction of mosquitoes at the 3rd and 4th weeks, respectively.