Edwards, S. L. et al. (2019) International Worm Meeting "Copious longevity via increased translation and proteasome activity."

Detienne, G., Cockx, B., Van De Walle, P., De Haes, W., Braeckman, B., Edwards, S. L., Temmerman, L., Schoofs, L.

[International Worm Meeting, 2019]

Royalactin, a glycoprotein thought to be essential for the development of long-lived queen honeybees, is capable of increasing longevity in several organisms. We previously showed that supplementing Caenorhabditis elegans with royalactin activates epidermal growth factor signaling and leads to an increased lifespan and improved health. Whilst many interventions that increase the lifespan of an organism rely on defensive strategies, which reduce growth factor signaling and protein synthesis and translation, we have found through proteomics and functional genetic approaches that royalactin treatment leads to an increase in protein translation, which is required for this lifespan extension. Additionally, we observe that proteasome activity is also upregulated and that there are no obvious trade-offs between royalactin-induced longevity and fecundity or stress resistance. Our work suggests a novel 'copious' strategy for healthy aging, in which organisms increase breakdown of proteins and replace these by upregulating the synthesis of new proteins, keeping their proteome in a young and healthy state and thereby increasing resistance to stressors. Our current focus is to investigate how increased protein turnover contributes to the subsequent effect on stress-resistance and healthy aging.

Edwards S L et al. (2010) Neuronal Development, Synaptic Function and Behavior, Madison, WI "Kinesin Acts with JIP3 and Dynein to Restrict the Flow of Golgi and Endosomal Organelles into Axons"

Richmond J E, Miller K G, Phillips B C, Edwards S L, Yu S

[Neuronal Development, Synaptic Function and Behavior, Madison, WI, 2010]

Prior studies hypothesized that kinesin (KIF5) and its putative adaptor JIP3 promote the anterograde movement of organelles from neuronal somas to axons. Contradicting that hypothesis, we show that C. elegans axons lacking UNC-16 (JIP3) accumulate Golgi, endosomes, and lysosomes at levels up to 10-fold higher than wild type, whereas ER membranes are largely unchanged. Reducing the function of UNC-116 (KIF5) or dynein produces similar hyper-anterograde transport phenotypes. Kinesin exerts at least one retrograde function by inhibiting an anterograde function of dynein. Hyper-anterograde transport of synaptic vesicle proteins from cell somas disrupts synaptic vesicle biogenesis and contributes to a reduction in mature synaptic vesicles at unc-16 mutant synapses. Together, the data reveal a membrane trafficking system that unexpectedly uses kinesin and JIP3 in a retrograde role to restrict the flow of Golgi and endosomal membranes into axons. Our findings highlight the challenges that dual regulated secretory pathways impose on neurons and provide insights into motor neuron diseases caused by impaired microtubule motor function.

Ali Khan L et al. (1994) Worm Breeder's Gazette "cej-1 Encodes a Novel Protein With Poly-Threonine Motif"

Siddiqui SS, Fukushige T, Ali Khan L, Tsukita Sh, Tabish M, Itoh M

[Worm Breeder's Gazette, 1994]

cej-1 Encodes a Novel Protein with Poly-Threonine Motif M. L. A. Khanl, M. Tabish, T. Fukushigel1 S. Tsukita2, M. Itoh , Sh. Tsukita , and S. S. Siddiqui. (1): Lab. of Molecular Biology, Dept of Ecological Engg. Toyohashi Univ. Technology, Toyohashi 441, and (2). National Institute for Physiological Sciences, Okazaki 444, Japan.

Ellwood RA et al. (2022) Commun Biol "Sulfur amino acid supplementation displays therapeutic potential in a C. elegans model of Duchenne muscular dystrophy."

Slade L, Torregrossa R, Etheridge T, Piasecki M, Sudevan S, Ellwood RA, Szewczyk NJ, Lewis J, Whiteman M

[Commun Biol, 2022]

Mutations in the dystrophin gene cause Duchenne muscular dystrophy (DMD), a common muscle disease that manifests with muscle weakness, wasting, and degeneration. An emerging theme in DMD pathophysiology is an intramuscular deficit in the gasotransmitter hydrogen sulfide (H₂S). Here we show that the C. elegans DMD model displays reduced levels of H₂S and expression of genes required for sulfur metabolism. These reductions can be offset by increasing bioavailability of sulfur containing amino acids (L-methionine, L-homocysteine, L-cysteine, L-glutathione, and L-taurine), augmenting healthspan primarily via improved calcium regulation, mitochondrial structure and delayed muscle cell death. Additionally, we show distinct differences in preservation mechanisms between sulfur amino acid vs H₂S administration, despite similarities in required health-preserving pathways. Our results suggest that the H₂S deficit in DMD is likely caused by altered sulfur metabolism and that modulation of this pathway may improve DMD muscle health via multiple evolutionarily conserved mechanisms.

Hasegawa M et al. (2014) FEBS Lett "Protein arginine methyltransferase 7 has a novel homodimer-like structure formed by tandem repeats."

Fukamizu A, Shimizu T, Kim JD, Toma-Fukai S, Hasegawa M

[FEBS Lett, 2014]

Protein arginine methyltransferase 7 (PRMT7) is a member of a family of enzymes that catalyze the transfer of methyl groups from S-adenosyl-l-methionine to nitrogen atoms on arginine residues. Here, we describe the crystal structure of Caenorhabditis elegans PRMT7 in complex with its reaction product S-adenosyl-L-homocysteine. The structural data indicated that PRMT7 harbors two tandem repeated PRMT core domains that form a novel homodimer-like structure. S-adenosyl-L-homocysteine bound to the N-terminal catalytic site only; the C-terminal catalytic site is occupied by a loop that inhibits cofactor binding. Mutagenesis demonstrated that only the N-terminal catalytic site of PRMT7 is responsible for cofactor binding.

Phillips B C et al. (2010) Neuronal Development, Synaptic Function and Behavior, Madison, WI "Forward Genetic Analysis of the Synaptic Gs Pathway"

Edwards S L, Phillips B C, Miller K G

[Neuronal Development, Synaptic Function and Behavior, Madison, WI, 2010]

Neurons communicate with each other and with muscle cells at synapses. A long term goal of this lab is to bridge the gap between synaptic function and the control of behavior by determining how signal transduction pathways regulate the activity of synapses. Our studies focus on the neuromuscular synapses that control locomotion in C. elegans. Genetic studies have shown that the integrated activities of at least 4 major, conserved Gα pathways control synaptic activity to produce the C. elegans locomotion behavior. Within this network, the neuronal Gαs pathway is an especially critical, but poorly understood, link between synaptic function and behavior- driving both primordial synaptic functions, such as C. elegans locomotion, and higher synaptic functions such as sleep, learning, and memory. However, the key proteins through which the Gαs pathway acts to mediate these functions remain poorly understood or unidentified. To address this, we have undertaken forward genetic screens to isolate mutants that suppress the strong phenotypes of kin-2(ce179) mutants, containing hyperactivated Protein Kinase A. These mutants exhibit hyperactive locomotion, slow growth, and aldicarb hypersensitivity resulting from overactivation of the Gαs pathway in both muscle cells and neurons. In Screen A, we used the COPAS Biosort to plate 3 adult kin-2(ce179) (F1 progeny of EMS-mutagenized animals) per well on 24-well plates and screened for adult F2 mutants with improved growth and/ or sluggish locomotion. In Screen B, we plated 200 kin-2(ce179) L1s (F2 progeny of mutagenized animals) per well on 24-well plates containing 0.1 mM Aldicarb and screened for the presence of F3 and F4 progeny after 7d at room temperature. Of 57 mutations from the combined screens, 42 are alleles of previously identified genes of known function, including the Gα signaling pathway proteins RIC-8, EGL-10, and KIN-2 (intragenic suppressors); presynaptic calcium channel proteins UNC-2, UNC-36, and CALF-1; synaptic vesicle priming and fusion proteins UNC-13 and UNC-18; and ACh receptor subunits UNC-38 and UNC-29. However, we also recovered mutations in two novel, conserved proteins of unknown function. We used SNPs to map mutations in 11 other loci to defined intervals and are awaiting whole genome sequencing data to identify which proteins they have disrupted.

Li X et al. (2016) Pharmacogn Mag "Life Span and Motility Effects of Ethanolic Extracts from Sophora moorcroftiana Seeds on Caenorhabditis elegans."

Han J, Ma X, Li X, Zhu R, Cui R, Dong K

[Pharmacogn Mag, 2016]

BACKGROUND: Sophora moorcroftiana is an endemic shrub species with a great value in folk medicine in Tibet, China. In this study, relatively little is known about whether S. moorcroftiana is beneficial in animals' nervous system and life span or not. MATERIALS AND METHODS: To address this question, under survival normal temperature (25C), S. moorcroftiana seeds were extracted with 95% ethanol, and Caenorhabditis elegans were exposed to three different extract concentrations (100 mg/L, 200 mg/L, and 400 mg/mL) from S. moorcroftiana seeds. RESULTS: The 95% ethanolic extracts from S. moorcroftiana seeds could increase life span and slow aging-related increase in C. elegans and could not obviously influence the motility of C. elegans. CONCLUSION: Given these results by our experiment for life span and motility with 95% ethanolic extracts from S. moorcroftiana seeds in C. elegans, the question whether S. moorcroftiana acts as an anti-aging substance in vivo arises. SUMMARY: The 95% ethanolic extracts from S. moorcroftiana seeds have no effect on the life span in C. elegans when extract concentrations from S. moorcroftiana seeds <400 mg/LThe 400 mg/L 95% ethanolic extracts from S. moorcroftiana seeds could increase life span in C. elegansThe 95% ethanolic extracts from S. moorcroftiana seeds could not obviously influence the motility in C. elegans. Abbreviation used: S. moorcroftiana: Sophora moorcroftiana; C. elegan: Caenorhabditis elegan; E. coli OP50: Escherichia coli OP50; DMSO: Dimethyl sulfoxide.

Satou T et al. (2002) Vet Parasitol "Assay of nematocidal activity of isoquinoline alkaloids using third-stage larvae of Strongyloides ratti and S. venezuelensis."

Tada I, Satou T, Matsuhashi R, Koike K, Koga M, Nikaido T

[Vet Parasitol, 2002]

We examined the effects of isoquinoline alkaloids in vitro in an effort to identify a treatment for Strongyloides stercoralis larva migrans in humans. Infective third-stage larvae of S. ratti and S. venezuelensis were used as model nematodes for S. stercoralis. Nematocidal activity was evaluated by the 50% paralysis concentration (PC(50)). Most of the tested isoquinoline alkaloids had activity for S. ratti and S. venezuelensis. We then evaluated in vitro cytotoxicity, which was the 50% inhibition concentration (IC(50)) of the compounds using HL60 tissue-culture cells. Three of the compounds (protopine, D-corydaline, and L-stylopine) which exhibited strong nematocidal activity, showed little cytotoxicity. In addition, we examined the relationship between nematocidal activity and cytotoxicity using the PC(50)/IC(50) ratio. A ratio equivalent to or lower than that calculated for the currently prescribed strongyloidosis treatments, ivermectin, albendazole and thiabendazole, was observed for allocryptopine, protopine, dehydrocorydaline, D-corydaline, L-stylopine, and papaverine. In contrast, the PC(50)/IC(50) ratios for protopine, D-corydaline, and L-stylopine were substantially more favorable. Therefore, protopine, D-corydaline, and L-stylopine were identified as potential effective treatments for strongyloidosis.

Petriv OI et al. (2004) J Mol Biol "A new definition for the consensus sequence of the peroxisome targeting signal type 2."

Tang L, Titorenko VI, Petriv OI, Rachubinski RA

[J Mol Biol, 2004]

All organisms except the nematode Caenorhabditis elegans have been shown to possess an import system for peroxisomal proteins containing a peroxisome targeting signal type 2 (PTS2). The currently accepted consensus sequence for this amino-terminal nonapeptide is -(R/K)(L/V/I)X-5(H/Q)(L/A)-. Some C. elegans proteins contain putative PTS2 motifs, including the ortholog (CeMeK) of human mevalonate kinase, an enzyme known to be targeted by PTS2 to mammalian peroxisomes. We cloned the gene for CeMeK (open reading frame Y42G9A.4) and examined the subcellular localization of CeMeK and of two other proteins with putative PTS2s at their amino termini encoded by the open reading frames D1053.2 and W10G11.11. All three proteins localized to the cytosol, confirming and extending the finding that C. elegans lacks PTS2-dependent peroxisomal protein import. The putative PTS2s of the proteins encoded by D1053.2 and W10G11.11 did not function in targeting to peroxisomes in yeast or mammalian cells, suggesting that the current PTS2 consensus sequence is too broad. Analysis of available experimental data on both functional and nonfunctional PTS2s led to two re-evaluated PTS2 consensus sequences: -R(L/V/I/Q)XX(L/V/I/H)(L/S/G/A)X(H/Q)(L/A)-, describes the most common variants of PTS2, while -(R/K)(L/V/I/Q)XX(L/V/I/H/ Q)(L/S/G/A/K)X(H/Q)(L/A/F)-, describes essentially all variants of PTS2. These redefined PTS2 consensus sequences will facilitate the identification of proteins of unknown cellular localization as possible peroxisomal proteins.

Oleh I Petriv et al. (2003) International Worm Meeting "Redefining the consensus sequence of the peroxisomal targeting signal type 2"

Vladimir I Titorenko, Richard A Rachubinski, Oleh I Petriv

[International Worm Meeting, 2003]

All organisms studied so far, except for the nematode Caenorhabditis elegans, have been shown to have developed an import system for peroxisomal proteins containing a peroxisomal targeting signal type 2 (PTS2). The currently accepted consensus sequence of this amino-terminal nonapeptide is (R/K)(L/V/I)X5(H/Q)(L/A). Some C. elegans proteins contain putative PTS2 motifs, including the ortholog (CeMeK) of human mevalonate kinase, an enzyme known to be targeted by a PTS2 to mammalian peroxisomes. We cloned the gene for CeMeK (Y42G9A.4) and examined the subcellular localization of CeMeK and of two other proteins with putative PTS2s at their amino termini encoded by the ORFs D1053.2 and W10G11.11. Two proteins (CeMeK and the product of ORF W10G11.11) localized to the cytosol, while the protein encoded by D1053.2 was found in nucleoli. The three putative PTS2s did not function in targeting to peroxisomes in yeast or mammalian cells, suggesting that the current PTS2 consensus sequence is too broad. After extensive analysis of available experimental data on both functional and nonfunctional PTS2s, we propose two re-evaluated PTS2 consensus sequences. The first, R(L/V/I)XX(L/V/I)(S/A/L/K)X(H/Q)(L/A), describes the most common PTS2 variants, while the second (R/K)(L/V/I/Q)XX(L/V/I/H/Q)(S/A/L/K/G)X(H/Q)(L/A/F) includes all rare variants of PTS2. Our study also confirms the finding that C. elegans apparently lacks any PTS2-dependant peroxisomal protein import.