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[
J Neurosci,
2003]
Thermotactic behavior in Caenorhabditis elegans is sensitive to both a worm's ambient temperature (T-amb) and its memory of the temperature of its cultivation (T-cult). The AFD neuron is part of a neural circuit that underlies thermotactic behavior. By monitoring the fluorescence of pH-sensitive green fluorescent protein localized to synaptic vesicles, we measured the rate of the synaptic release of AFD in worms cultivated at temperatures between 15 and 25degreesC, and subjected to fixed, ambient temperatures in the same range. We found that the rate of AFD synaptic release is high if either T-amb > T-cult or T-amb > T-cult, but AFD synaptic release is low if T-amb congruent to T-cult. This suggests that AFD encodes a direct comparison between T-amb and T-cult.
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[
Trends Mol Med,
2007]
Transforming growth factor beta1 (TGFbeta1), an important pleiotropic, immunoregulatory cytokine, uses distinct signaling mechanisms in lymphocytes to affect T-cell homeostasis, regulatory T (T(reg))-cell and effector-cell function and tumorigenesis. Defects in TGFbeta1 expression or its signaling in T cells correlate with the onset of several autoimmune diseases. TGFbeta1 prevents abnormal T-cell activation through the modulation of Ca(2+)-calcineurin signaling in a Caenorhabditis elegans Sma and Drosophila Mad proteins (SMAD)3 and SMAD4-independent manner; however, in T(reg) cells, its effects are mediated, at least in part, through SMAD signaling. TGFbeta1 also acts as a pro-inflammatory cytokine and induces interleukin (IL)-17-producing pathogenic T-helper cells (T(h) IL-17 cells) synergistically during an inflammatory response in which IL-6 is produced. Here, we will review TGFbeta1 and its signaling in T cells with an emphasis on the regulatory arm of immune tolerance.
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[
Genomics,
1995]
Recently, a novel family of genes with a region of homology to the mouse T locus, which is known to play a crucial, and conserved, role in vertebrate development, has been discovered. The region of homology has been named the T-box. The T-box domain of the prototypical T locus product is associated with sequence-specific DNA binding activity. In this report, we have characterized four members of the T-box gene family from the nematode Caenorhabditis elegans. All lie in close proximity to each other in the middle of chromosome III. Homology analysis among all completely sequenced T-box products indicates a larger size for the conserved T-box domain (166 to 203 residues) than previously reported. Phylogenetic analysis suggests that one C. elegans T-box gene may be a direct ortholog of the mouse Tbx2 and Drosophila omb genes. The accumulated data demonstrate the ancient nature of the T-box gene family and suggest the existence of at least three separate T-box-containing genes in a common early metazoan ancestor to nematodes and vertebrates.
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[
BMC Mol Cell Biol,
2022]
BACKGROUND: Protein-protein interactions form the basis of every organism and thus, investigating their dynamics, intracellular protein localization, trafficking and interactions of distinct proteins such as receptors and their ligand-binding are of general interest. Bioluminescence resonance energy transfer (BRET) is a powerful tool to investigate these aspects in vitro. Since in vitro approaches mostly neglect the more complex in vivo situation, we established BRET as an in vivo tool for studying protein interactions in the nematode C. elegans. RESULTS: We generated worms expressing NanoBRET sensors and elucidated the interaction of two ligand-G protein-coupled receptor (GPCR) pairs, the neuropeptide receptor NPR-11 and the Adhesion GPCR LAT-1. Furthermore, we adapted the enhanced bystander BRET technology to measure subcellular protein localization. Using this approach, we traced ligand-induced internalization of NPR-11 in vivo. CONCLUSIONS: Our results indicate that in vivo NanoBRET is a tool to investigate specific protein interactions and localization in a physiological setting in real time in the living organism C. elegans.
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[
Glycobiology,
2006]
The common O-glycan core structure in animal glycoproteins is the core 1 disaccharide Galbeta1-3GalNAcalpha1-Ser/Thr, which is generated by addition of Gal to GalNAcalpha1-Ser/Thr by core 1 UDP-Gal:GalNAcalpha1-Ser/Thr beta1,3-galactosyltransferase (core 1 beta3-Gal-T or T-synthase, EC2.4.1.122)(2). Although O-glycans play important roles in vertebrates, much remains to be learned from model organisms such as the free-living nematode Caenorhabditis elegans, which offer many advantages in exploring O-glycan structure/function. Here we report the cloning and enzymatic characterization of T-synthase from C. elegans (Ce-T-synthase). A putative C. elegans gene for T-synthase, C38H2.2, was identified in GenBank by a BlastP search using the human T-synthase protein sequence. The full-length cDNA for Ce-T-synthase, which was generated by PCR using a C. elegans cDNA library as the template, contains 1,170 bp including the stop TAA. The cDNA encodes a protein of 389 amino acids with typical type-II membrane topology and a remarkable 42.7% identity to the human T-synthase. Ce-T-synthase has 7 Cys residues in the lumenal domain including 6 conserved Cys residues in all of the orthologs. The Ce-T-synthase has 4 potential N-glycosylation sequons, whereas the mammalian orthologs lack N-glycosylation sequons. Only one gene for Ce-T-synthase was identified in the genome-wide search and it contains 8 exons. Promoter analysis of the Ce-T-synthase using green fluorescent protein constructs show that the gene is expressed at all developmental stages and appears to be in all cells. Unexpectedly, only minimal activity was recovered in the recombinant, soluble Ce-T-synthase secreted from a wide variety of mammalian cell lines, whereas robust enzyme activity was recovered in the soluble Ce-T-synthase expressed in Hi-5 insect cells. Vertebrate T-synthase requires the molecular chaperone Cosmc, but our results show that Ce-T-synthase does not require Cosmc, and might require invertebrate-specific factors for formation of the optimally active enzyme. These results show that the Ce-T-synthase is a functional ortholog to the human T-synthase in generating core 1 O-glycans and opens new avenues to explore O-glycan function in this model organism.
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[
International Worm Meeting,
2019]
Orientation of spindles and cell division planes is vital for tissue formation and cellular communication in many species. Especially during embryogenesis it ensures that correct cell-cell contacts are established, correct shapes are generated and an organism can develop. Although several signaling pathways in oriented cell division have been well characterised such as wnt/frizzled, there is strong evidence for additional signal pathways especially in controlling early anterior-posterior polarity decisions. Recently, we have identified the homolog of the Adhesion G protein-coupled receptor Latrophilin, LAT-1, as a novel player in oriented cell division in the early C. elegans embryo. The cell surface receptor controls the proper anterior-posterior direction of cell division of specific blastomeres by a classical G protein-cascade based on coupling of the receptor to a Gs protein (GSA-1) which leads to elevated intracellular levels of the second messenger cyclic AMP (cAMP). Thus, strikingly, a metabotropic signal controls distinct aspects of polarity. We are combining in vitro and in vivo approaches to elucidate how LAT-1 mediates anterior-posterior cell division plane orientation via a non-polarised signal and by which mechanisms polarity information is elicited. We identified EPAC-1 to be the downstream effector of cAMP by testing specific effector compounds for their ability to generate a phenocopy of a
lat-1 null mutant which displays defective embryonic cell division plane orientations in ABal descendants. To better elucidate the role of the second messager we employed two different tools. We used a photoactivable adenylyl cyclase to mimic the LAT-1 signal at different stages of embryogenesis to gain a comprehensive picture of the spatial and temporal requirements and effects of LAT-1 within the developing embryo. Further, a cAMP biosensor to follow cAMP generation and distribution upon LAT-1 pathway stimulation by FRET analysis. Taken together, our data contribute to address the question how a metabotropic signal can mediate a polarised process.
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[
Int J Syst Evol Microbiol,
2007]
A yellow-pigmented, Gram-positive, aerobic, non-motile, non-spore-forming, irregular rod-shaped bacterium (strain TAN 31504(T)) was isolated from the bacteriophagous nematode Caenorhabditis elegans. Based on 16S rRNA gene sequence similarity, DNA G+C content of 69.5 mol%, 2,4-diaminobutyric acid in the cell-wall peptidoglycan, major menaquinone MK-11, abundance of anteiso- and iso-fatty acids, polar lipids diphosphatidylglycerol and phosphatidylglycerol and a number of shared biochemical characteristics, strain TAN 31504(T) was placed in the genus Leucobacter. DNA-DNA hybridization comparisons demonstrated a 91 % DNA-DNA relatedness between strain TAN 31504(T) and Leucobacter chromiireducens LMG 22506(T) indicating that these two strains belong to the same species, when the recommended threshold value of 70 % DNA-DNA relatedness for the definition of a bacterial species by the ad hoc committee on reconciliation of approaches to bacterial systematics is considered. Based on distinct differences in morphology, physiology, chemotaxonomic markers and various biochemical characteristics, it is proposed to split the species L. chromiireducens into two novel subspecies, Leucobacter chromiireducens subsp. chromiireducens subsp. nov. (type strain L-1(T)=CIP 108389(T)=LMG 22506(T)) and Leucobacter chromiireducens subsp. solipictus subsp. nov. (type strain TAN 31504(T)=DSM 18340(T)=ATCC BAA-1336(T)).
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[
International Worm Meeting,
2019]
The neuropeptide Y (NPY) family is a well-studied system due to the essential roles in regulating food consumption, control of mood and anxiety or ethanol intake. Several neuropeptides activate different G protein-coupled receptors (GPCRs), thus forming a multireceptor/ multiligand system, making neuropeptidergic signaling highly complex. Being a key regulator of essential processes, it is not surprising that neuropeptide signaling is conserved down to basal animals such as protostomes and thus, forms an ancient system. Surprisingly, in Caenorhabditis elegans, there is controversy on the existence of an NPY orthologous system. Upon the discovery of the FMRFamide-like peptide (FLP)/ NPY/ RFamide-like receptor (NPR) system in the nematode, orthology to the human system was suggested. However, later global phylogenetic studies indicated that FLP/ NPR is not truly homologous to the NPY system, but nematode-specific. To gain insights in the pharmacological and functional similarities of the NPR/ FLP and the human NPY system, we conducted a comprehensive comparative pharmacological study of the FLP/ NPR system proving that G protein-coupling and ligand-binding modes are similar to the human NPY system. Further, in vitro and in vivo analyses show cross-reactivity of the NPY with the FLP/ NPR system culminating in the ability of the human GPCRs to functionally substitute FLP/ NPR signaling in C. elegans in vivo. This functional homology of both neuropeptide systems was subsequently utilized to identify FLP-14 as a main driver for chemotaxis. Taken together, our data reveal high levels of pharmacological and functional similarity of human and C. elegans NPY systems, suggesting a homologous relationship, and highlight the importance for physiological as well as molecular studies. Ultimately, our study will contribute to the understanding on how specific neuropeptide receptor signals are translated into physiological function.
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[
Genome,
1997]
The T-box gene family consists of members that share a unique DNA binding domain. The best characterized T-box gene, Brachyury or T, encodes a transcription factor that plays an important role in early vertebrate development. Seven other recently described mouse T-box genes are also expressed during development. In the nematode Caenorhabditis elegans, four T-box genes have been characterized to date. In this study, we describe three new C. elegans T-box genes, named
Ce-tbx-11,
Ce-tbx-12, and
Ce-tbx-17.
Ce-tbx-11 and
Ce-tbx-17 were uncovered through the sequencing efforts of the C. elegans Genome Project.
Ce-tbx-12 was uncovered through degenerate PCR analysis of C. elegans genomic DNA.
Ce-tbx-11 and
Ce-tbx-17 are located in close proximity to the four other previously described T-box genes in the central region of chromosome III. In contrast,
Ce-tbx-12 maps alone to chromosome II. Phylogenetic analysis of all known T-box domain sequences provides evidence of an ancient origin for this gene family.
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[
European Worm Meeting,
2002]
T-box genes are a group of developmentally important transcription factors united by a common DNA binding domain. T-box genes are present in all metazoan species so far analysed but are absent from yeast. There are 20 T-box genes in C. elegans, more than twice the number found in Drosophila. Many of the C. elegans T-box genes are highly diverged from those found in other species while others have clear orthologues present throughout the metazoan kingdom. One highly conserved T-box gene is
mab-9, a member of the
tbx20 sub-family1. This was the first C. elegans T-box gene to be identified by mutation and is required for cell fate specification during hindgut and male tail development, and aspects of nervous system function. One other conserved T-box gene has recently been reported to be important for a particular muscle cell fate specification2. We have inactivated the remaining C. elegans T-box genes by RNAi and have found obvious phenotypes only in very few cases. These phenotypes include embryonic lethality, L1 lethality, and a Dpy phenotype with weakly penetrant male tail defects, and will be described in detail. The remaining T-box genes give no obvious phenotype by RNAi. Phylogenetic analysis reveals that several pairs of T-box genes are very similar to eachother and are therefore likely to be the result of recent duplications. This might suggest functional redundancy. Double RNAi experiments have revealed this to be the case with at least two of the T-box gene pairs (see also poster by Pocock et al). Study of the expression patterns of the whole T-box family may suggest other potential redundancy relationships which can be explored by RNAi. Comparison of the C. elegans T-box genes with the set of T-box genes now defined for C. briggsae is being used as a tool for defining potentially important regulatory regions present in orthologous genes.