scholarly journals Pkc-Mediated Stimulation of Amphibian Cftr Depends on a Single Phosphorylation Consensus Site. Insertion of This Site Confers Pkc Sensitivity to Human Cftr

2001 ◽  
Vol 117 (5) ◽  
pp. 457-468 ◽  
Author(s):  
Brian Button ◽  
Luis Reuss ◽  
Guillermo A. Altenberg
Keyword(s):  
Critical Role ◽  
Phosphorylation Site ◽  
Minor Effect ◽  
Phosphorylation Sites ◽  
Net Charge ◽  
Necturus Maculosus ◽  
Equivalent Site ◽  
A Minor ◽  
Stimulation Of ◽  
R Domain

Mutations of the CFTR, a phosphorylation-regulated Cl− channel, cause cystic fibrosis. Activation of CFTR by PKA stimulation appears to be mediated by a complex interaction between several consensus phosphorylation sites in the regulatory domain (R domain). None of these sites has a critical role in this process. Here, we show that although endogenous phosphorylation by PKC is required for the effect of PKA on CFTR, stimulation of PKC by itself has only a minor effect on human CFTR. In contrast, CFTR from the amphibians Necturus maculosus and Xenopus laevis (XCFTR) can be activated to similar degrees by stimulation of either PKA or PKC. Furthermore, the activation of XCFTR by PKC is independent of the net charge of the R domain, and mutagenesis experiments indicate that a single site (Thr665) is required for the activation of XCFTR. Human CFTR lacks the PKC phosphorylation consensus site that includes Thr665, but insertion of an equivalent site results in a large activation upon PKC stimulation. These observations establish the presence of a novel mechanism of activation of CFTR by phosphorylation of the R domain, i.e., activation by PKC requires a single consensus phosphorylation site and is unrelated to the net charge of the R domain.

scholarly journals Coregulation of dimorphism and symbiosis by cyclic AMP signaling in the lichenized fungusUmbilicaria muhlenbergii

2020 ◽  
Vol 117 (38) ◽  
pp. 23847-23858
Author(s):  
Yanyan Wang ◽  
Xinli Wei ◽  
Zhuyun Bian ◽  
Jiangchun Wei ◽  
Jin-Rong Xu
Keyword(s):  
Close Contact ◽  
Critical Role ◽  
Hyphal Growth ◽  
Hyperosmotic Stress ◽  
Yeast Cells ◽  
Minor Effect ◽  
Symbiotic Interaction ◽  
Pseudohyphal Growth ◽  
Lichenized Fungus ◽  
A Minor

Umbilicaria muhlenbergiiis the only known dimorphic lichenized fungus that grows in the hyphal form in lichen thalli but as yeast cells in axenic cultures. However, the regulation of yeast-to-hypha transition and its relationship to the establishment of symbiosis are not clear. In this study, we show that nutrient limitation and hyperosmotic stress trigger the dimorphic change inU. muhlenbergii. Contact with algal cells of its photobiontTrebouxia jamesiiinduced pseudohyphal growth. Treatments with the cAMP diphosphoesterase inhibitor IBMX (3-isobutyl-1-methylxanthine) induced pseudohyphal/hyphal growth and resulted in the differentiation of heavily melanized, lichen cortex-like structures in culture, indicating the role of cAMP signaling in regulating dimorphism. To confirm this observation, we identified and characterized two Gα subunitsUmGPA2andUmGPA3. Whereas deletion ofUmGPA2had only a minor effect on pseudohyphal growth, the ΔUmgpa3mutant was defective in yeast-to-pseudohypha transition induced by hyperosmotic stress orT. jamesiicells. IBMX treatment suppressed the defect of ΔUmgpa3in pseudohyphal growth. Transformants expressing theUmGPA3G45VorUmGPA3Q208Ldominant active allele were enhanced in the yeast-to-pseudohypha transition and developed pseudohyphae under conditions noninducible to the wild type. Interestingly,T. jamesiicells in close contact with pseudohyphae ofUmGPA3G45VandUmGPA3Q208Ltransformants often collapsed and died after coincubation for over 72 h, indicating that improperly regulated pseudohyphal growth due to dominant active mutations may disrupt the initial establishment of symbiotic interaction between the photobiont and mycobiont. Taken together, these results show that the cAMP-PKA pathway plays a critical role in regulating dimorphism and symbiosis inU. muhlenbergii.

Regulation of the segmental swim-generating system by a pair of identified interneurons in the leech head ganglion

1995 ◽  
Vol 73 (3) ◽  
pp. 983-992 ◽  
Author(s):  
P. D. Brodfuehrer ◽  
H. J. Parker ◽  
A. Burns ◽  
M. Berg
Keyword(s):  
Single Cell ◽  
Motor Neurons ◽  
Synaptic Input ◽  
Dorsal Surface ◽  
Firing Frequency ◽  
Activity Level ◽  
Minor Effect ◽  
Excitatory Synaptic Input ◽  
A Minor ◽  
Stimulation Of

1. The aim of this study was to identify neurons that modulate activity of segmental swim gating interneurons. We found a pair of bilaterally symmetrical interneurons, cells SE1, whose activity level directly influences three groups of segmental neurons associated with generating swimming in the medicinal leech. 2. The somata of cells SE1 are located on the dorsal surface of the subesophageal ganglion. Their axons extend most, if not the entire, length of the ventral nerve cord and appear to make identical connections with the same group of swim-generating neurons in all segmental ganglia. 3. Cells SE1 excite monosynaptically all segmental swim gating interneurons, cells 204, examined. The level of excitation in cell 204 is directly correlated with the firing frequency of cell SE1. In most quiescent preparations (when the preparation is not swimming) hyperpolarization of a single cell SE1 eliminates all excitatory synaptic input to cells 204. 4. Cells SE1 excite monosynaptically three swim oscillatory interneurons, cells 115, 28, and 208. The strength of the connection from cell SE1 to cell 115 is stronger than the connection from cell SE1 to either cells 28 or 208. The level of excitation in cell 115 is directly correlated with the firing frequency of cell SE1. In most quiescent preparations, hyperpolarization of a single cell SE1 eliminates all excitatory synaptic input to cell 115 but has only a minor effect on the level of activity in cells 208 and 28. 5. Due most likely to the strong and direct connections cells SE1 have with swim gating and oscillatory interneurons, brief stimulation of cell SE1 can elicit swimming. Swimming generally occurs within 1 s after stimulation of cell SE1. During swimming, the membrane potential of cell SE1 depolarizes by 2-5 mV, and its firing frequency increases. Brief depolarization of cell SE1 during swimming reliably shifts the phase of the swimming rhythm, whereas longer periods of depolarization increase both swim period and burst duration. 6. Excitatory motor neurons to the dorsal longitudinal muscles, cells 3, 5, and 7, are strongly excited by stimulation of cell SE1. The firing frequency of cell 3 is positively correlated with the firing frequency of cell SE1. 7. The results of this study indicate that cells SE1 can modulate the level of excitation in three groups of neurons associated with generating leech swimming.(ABSTRACT TRUNCATED AT 400 WORDS)

ACh release from horse airway cholinergic nerves: effects of stimulation intensity and muscle preload

1993 ◽  
Vol 264 (3) ◽  
pp. L269-L275 ◽  
Author(s):  
Z. Wang ◽  
N. E. Robinson ◽  
M. Yu
Keyword(s):  
Release Rate ◽  
Electrical Field Stimulation ◽  
Minor Effect ◽  
Field Stimulation ◽  
Cholinergic Nerves ◽  
Ach Release ◽  
Electrical Field ◽  
Stimulation Parameters ◽  
A Minor ◽  
Stimulation Of

This study was conducted to determine the effects of stimulation parameters and muscle preload on acetylcholine (ACh) release induced by electrical field stimulation (EFS) of horse airway cholinergic nerves. Trachealis strip bundles were prepared and suspended in 2-ml tissue baths. The tissues were stimulated three to five times for 30 min each. Increasing frequency (0.5-16 Hz) and voltage (5-20 V) increased ACh release; increasing pulse duration (0.5-3 ms) had only a minor effect. Alterations in muscle preload (2-20 g) had no effect on ACh release. ACh release was fairly constant for up to five repeated stimulation periods with the same EFS parameters. Stimulation of the tissues for 15 min released the same amount of ACh as 30 min if the amount was expressed as picomoles per gram per minute, suggesting that ACh release rate was constant during the 30-min period of stimulation. Atropine (10(-6) M) potentiated the release of ACh four- to fivefold, presumably by removing the autoinhibitory effect of ACh on the cholinergic nerves. Tetrodotoxin (10(-6) M) abolished the EFS-induced ACh release.

scholarly journals Proteolytic cleavage and PKA phosphorylation of α1C subunit are not required for adrenergic regulation of CaV1.2 in the heart

2017 ◽  
Vol 114 (34) ◽  
pp. 9194-9199 ◽  
Author(s):  
Alexander Katchman ◽  
Lin Yang ◽  
Sergey I. Zakharov ◽  
Jared Kushner ◽  
Jeffrey Abrams ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Molecular Mechanisms ◽  
Calcium Influx ◽  
Phosphorylation Site ◽  
Proteolytic Cleavage ◽  
Phosphorylation Sites ◽  
Adrenergic Stimulation ◽  
Adrenergic Regulation ◽  
Voltage Dependent ◽  
Stimulation Of

Calcium influx through the voltage-dependent L-type calcium channel (CaV1.2) rapidly increases in the heart during “fight or flight” through activation of the β-adrenergic and protein kinase A (PKA) signaling pathway. The precise molecular mechanisms of β-adrenergic activation of cardiac CaV1.2, however, are incompletely known, but are presumed to require phosphorylation of residues in α1C and C-terminal proteolytic cleavage of the α1C subunit. We generated transgenic mice expressing an α1C with alanine substitutions of all conserved serine or threonine, which is predicted to be a potential PKA phosphorylation site by at least one prediction tool, while sparing the residues previously shown to be phosphorylated but shown individually not to be required for β-adrenergic regulation of CaV1.2 current (17-mutant). A second line included these 17 putative sites plus the five previously identified phosphoregulatory sites (22-mutant), thus allowing us to query whether regulation requires their contribution in combination. We determined that acute β-adrenergic regulation does not require any combination of potential PKA phosphorylation sites conserved in human, guinea pig, rabbit, rat, and mouse α1C subunits. We separately generated transgenic mice with inducible expression of proteolytic-resistant α1C. Prevention of C-terminal cleavage did not alter β-adrenergic stimulation of CaV1.2 in the heart. These studies definitively rule out a role for all conserved consensus PKA phosphorylation sites in α1C in β-adrenergic stimulation of CaV1.2, and show that phosphoregulatory sites on α1C are not redundant and do not each fractionally contribute to the net stimulatory effect of β-adrenergic stimulation. Further, proteolytic cleavage of α1C is not required for β-adrenergic stimulation of CaV1.2.

Improvement of monitoring of tertiary filtration with particle counting

2006 ◽  
Vol 6 (1) ◽  
pp. 1-9
Author(s):  
V. Miska ◽  
J.H.J.M. van der Graaf ◽  
J. de Koning
Keyword(s):  
Particle Surface ◽  
Minor Effect ◽  
Particle Size Distributions ◽  
Particle Volume ◽  
Mass Distributions ◽  
Particle Counting ◽  
Total Particle ◽  
Particle Counts ◽  
A Minor ◽  
Sample Dilution

Nowadays filtration processes are still monitored with conventional analyses like turbidity measurements and, in case of flocculation–filtration, with phosphorus analyses. Turbidity measurements have the disadvantage that breakthrough of small flocs cannot be displayed, because of the blindness regarding changes in the mass distributions. Additional particle volume distributions calculated from particle size distributions (PSDs) would provide a better assessment of filtration performance. Lab-scale experiments have been executed on a flocculation–filtration column fed with effluent from WWTP Beverwijk in The Netherlands. Besides particle counting at various sampling points, the effect of sample dilution on the accuracy of PSD measurements has been reflected. It was found that the dilution has a minor effect on PSD of low turbidity samples such as process filtrate. The correlation between total particle counts, total particle volume (TPV) and total particle surface is not high but is at least better for diluted measurements of particles in the range 2–10 μm. Furthermore, possible relations between floc-bound phosphorus and TPV removal had been investigated. A good correlation coefficient is found for TPV removal versus floc-bound phosphorus removal for the experiments with polyaluminiumchloride and the experiments with single denitrifying and blank filtration.

scholarly journals Linkage mapping identifies a non-synonymous mutation in FLOWERING LOCUS T (FT-B1) increasing spikelet number per spike

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jonathan Brassac ◽  
Quddoos H. Muqaddasi ◽  
Jörg Plieske ◽  
Martin W. Ganal ◽  
Marion S. Röder
Keyword(s):  
Flowering Time ◽  
Aspartic Acid ◽  
Synonymous Substitution ◽  
Minor Effect ◽  
Phenotypic Variance ◽  
Spikelet Number ◽  
Wheat Varieties ◽  
Trait Locus ◽  
A Minor ◽  
Spikelet Number Per Spike

AbstractTotal spikelet number per spike (TSN) is a major component of spike architecture in wheat (Triticumaestivum L.). A major and consistent quantitative trait locus (QTL) was discovered for TSN in a doubled haploid spring wheat population grown in the field over 4 years. The QTL on chromosome 7B explained up to 20.5% of phenotypic variance. In its physical interval (7B: 6.37–21.67 Mb), the gene FLOWERINGLOCUST (FT-B1) emerged as candidate for the observed effect. In one of the parental lines, FT-B1 carried a non-synonymous substitution on position 19 of the coding sequence. This mutation modifying an aspartic acid (D) into a histidine (H) occurred in a highly conserved position. The mutation was observed with a frequency of ca. 68% in a set of 135 hexaploid wheat varieties and landraces, while it was not found in other plant species. FT-B1 only showed a minor effect on heading and flowering time (FT) which were dominated by a major QTL on chromosome 5A caused by segregation of the vernalization gene VRN-A1. Individuals carrying the FT-B1 allele with amino acid histidine had, on average, a higher number of spikelets (15.1) than individuals with the aspartic acid allele (14.3) independent of their VRN-A1 allele. We show that the effect of TSN is not mainly related to flowering time; however, the duration of pre-anthesis phases may play a major role.

scholarly journals A deep learning based approach for prediction of Chlamydomonas reinhardtii phosphorylation sites

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Niraj Thapa ◽  
Meenal Chaudhari ◽  
Anthony A. Iannetta ◽  
Clarence White ◽  
Kaushik Roy ◽  
...  
Keyword(s):  
Deep Learning ◽  
Chlamydomonas Reinhardtii ◽  
Protein Phosphorylation ◽  
Short Term Memory ◽  
Phosphorylation Site ◽  
Specific Protein ◽  
Training Dataset ◽  
Phosphorylation Sites ◽  
Site Prediction ◽  
Model Combining

AbstractProtein phosphorylation, which is one of the most important post-translational modifications (PTMs), is involved in regulating myriad cellular processes. Herein, we present a novel deep learning based approach for organism-specific protein phosphorylation site prediction in Chlamydomonas reinhardtii, a model algal phototroph. An ensemble model combining convolutional neural networks and long short-term memory (LSTM) achieves the best performance in predicting phosphorylation sites in C. reinhardtii. Deemed Chlamy-EnPhosSite, the measured best AUC and MCC are 0.90 and 0.64 respectively for a combined dataset of serine (S) and threonine (T) in independent testing higher than those measures for other predictors. When applied to the entire C. reinhardtii proteome (totaling 1,809,304 S and T sites), Chlamy-EnPhosSite yielded 499,411 phosphorylated sites with a cut-off value of 0.5 and 237,949 phosphorylated sites with a cut-off value of 0.7. These predictions were compared to an experimental dataset of phosphosites identified by liquid chromatography-tandem mass spectrometry (LC–MS/MS) in a blinded study and approximately 89.69% of 2,663 C. reinhardtii S and T phosphorylation sites were successfully predicted by Chlamy-EnPhosSite at a probability cut-off of 0.5 and 76.83% of sites were successfully identified at a more stringent 0.7 cut-off. Interestingly, Chlamy-EnPhosSite also successfully predicted experimentally confirmed phosphorylation sites in a protein sequence (e.g., RPS6 S245) which did not appear in the training dataset, highlighting prediction accuracy and the power of leveraging predictions to identify biologically relevant PTM sites. These results demonstrate that our method represents a robust and complementary technique for high-throughput phosphorylation site prediction in C. reinhardtii. It has potential to serve as a useful tool to the community. Chlamy-EnPhosSite will contribute to the understanding of how protein phosphorylation influences various biological processes in this important model microalga.

scholarly journals IL-6 Is Not Absolutely Essential for the Development of a TH17 Immune Response after an Aerosol Infection with Mycobacterium tuberculosis H37rv

Cells ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 9
Author(s):  
Kristina Ritter ◽  
Jan Christian Sodenkamp ◽  
Alexandra Hölscher ◽  
Jochen Behrends ◽  
Christoph Hölscher
Keyword(s):  
Immune Response ◽  
Inflammatory Diseases ◽  
Mycobacterial Infection ◽  
Minor Effect ◽  
Mycobacterium Tuberculosis H37rv ◽  
Th 17 ◽  
Anti Inflammatory ◽  
A Minor ◽  
Aerosol Infection ◽  
Deficient Mice

Anti-inflammatory treatment of chronic inflammatory diseases often increases susceptibility to infectious diseases such as tuberculosis (TB). Since numerous chronic inflammatory and autoimmune diseases are mediated by interleukin (IL)-6-induced T helper (TH) 17 cells, a TH17-directed anti-inflammatory therapy may be preferable to an IL-12-dependent TH1 inhibition in order to avoid reactivation of latent infections. To assess, however, the risk of inhibition of IL-6-dependent TH17-mediated inflammation, we examined the TH17 immune response and the course of experimental TB in IL-6- and T-cell-specific gp130-deficient mice. Our study revealed that the absence of IL-6 or gp130 on T cells has only a minor effect on the development of antigen-specific TH1 and TH17 cells. Importantly, these gene-deficient mice were as capable as wild type mice to control mycobacterial infection. Together, in contrast to its key function for TH17 development in other inflammatory diseases, IL-6 plays an inferior role for the generation of TH17 immune responses during experimental TB.

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