Michael Hoffmann et al. (2006) European Worm Meeting "The Planar Cell Polarity Protein VANG-1 interferes with Intercalation Events during Epithelial Morphogenesis In C. elegans"

Michael Hoffmann, Christoph Segbert, Olaf Bossinger, Gisela Helbig

[European Worm Meeting, 2006]

Michael Hoffmann, Christoph Segbert, Gisela Helbig and Olaf Bossinger. C. elegans VANG-1 is the only homolog of the Drosophila planar cell polarity (PCP) protein Strabismus/Van Gogh. We originally identified VANG-1 as a putative binding partner of DLG-1 (Discs large), a MAGUK protein that acts as a molecular scaffold for the establishment of the apical junction in all epithelia of the C. elegans embryo. The phenotype observed in vang-1 mutant embryos show intercalation defects during epithelial morphogenesis of the intestine and the hypodermis. Similar phenotypes arise in lin-17 (frizzled) mutants or after RNAi against dsh-2 (dishevelled), indicating core components of the PCP pathway in other systems to be involved in cell intercalation in the C. elegans embryo. In addition, VANG-1 and DSH-2 do colocalize and the asymmetric distribution of VANG-1 in epithelia depends on DSH-2. Taken together these results suggests that planar cell polarity might exist in C. elegans.

Peres, Tanara et al. (2021) International Worm Meeting "Nutrient-induced rewiring of microbial metabolic pathways modulate 5-fluorouracil efficacy in C. elegans"

Zimmermman, Johannes, Kaleta, Christoph, Chrysostomou, Despoina, Marchesi, Julian, W. Zatorska, Michalina, Barr, Alexis, Quintaneiro, Leonor, Cabreiro, Filipe, Martinez-Martinez, Daniel, Peres, Tanara, Kramer, Holger, Best, Lena, Marinos, Georgios, B. Mokochinski, Joao

[International Worm Meeting, 2021]

Microbiota derived metabolites have been shown to influence cancer susceptibility, tumour progression and therapy outcome. Diet is a major environmental factor influencing gut microbiota composition, dictating the abundance of potential oncometabolites. The complex interactions between diet and the microbiota need to be clarified so that dietary intake may be shaped to influence disease outcome. Having previously demonstrated the impact of bacterial metabolism in 5-fluorouracil (5-FU) cancer drug efficacy, we set out to investigate the role of nutrition in host-microbe responses to this drug, an antimetabolite widely used for colorectal cancer (CRC) chemotherapy. Host-microbe-drug-nutrient interaction and its influence on drug efficacy were assessed using a high-throughput 4-way screening approach. To understand gene-nutrient interactions at the bacterial level that regulate the effect of a drug on host physiology, genetically modified E. coli from the Keio deletion library were used to probe the role of bacterial metabolic pathways in 5-FU effects in a nutrient-dependent manner. We found that glycolytic nutrients, including sugars, importantly antagonise the toxic effects of 5-FU action on the host. Sugars induce a shift in metabolic pathways that favour the production of nucleotides, specifically UMP and UTP derived from the activation of pyrimidine de novo biosynthesis pathway in bacteria, counteracting pro-drug activation through the salvage pathway. We also found that bacterial mutations in the TCA cycle and pyruvate metabolism decrease drug efficacy in amino acid-based media. A combination of bacterial genetic work and metabolomics has revealed the presence of a novel bacterial metabolite with the capacity to increase 5-FU drug efficacy in both C. elegans and human cancer cells. Importantly, flux balance analysis of the cancer associated microbiota predicts an increased production of this metabolite in several organs. In particular, investigations of the gut microbiome from four independent human CRC cohorts shows an increased production of this metabolite, compared to healthy patients, further implicating this metabolite in drug cancer therapy. These findings highlight the potential of manipulating gut microbiota through diet to improve cancer therapy outcome.

Li, C.C.Y. et al. (2017) International Worm Meeting "Towards a unified global C. elegans Metabolic Reconstruction Model."

Li, C.C.Y., Witting, M., Kaleta, C., Casanueva, O, Hastings, J., Le Novere, N.

[International Worm Meeting, 2017]

C. elegans has recently been advanced as a premier metazoan model organism for the study of metabolism, with the publication of two whole-genome metabolic models (1, 2). Using these models together with -omics data allows the in-depth data-driven exploration of systems-level metabolism using in silico simulations. In a GENiE workshop to be held April 2017 at the Babraham Institute, Cambridge, UK, the relationships between these two existing metabolic models will be explored with the objective of generating a consensus model. Because the two reconstructions are still incomplete, and certain important pathways and areas of metabolism are currently under-annotated, we aim to identify specific areas that are relevant to the C. elegans community and prioritise them for further annotation in a follow-up community-driven "annotation jamboree" workshop. This poster will describe the main objectives set by the first workshop and opens the invitation to the C. elegans metabolic research community to contribute to the follow-up annotation efforts. 1. Gebauer, J.; Gentsch, C.; Mansfeld, J.; Schmei beta er, K.; Waschina, S.; Brandes, S.; Klimmasch, L.; Zamboni, N.; Zarse, K.; Schuster, S.; Ristow, M.; Schauble, S. & Kaleta, C. (2016), 'A Genome-Scale Database and Reconstruction of Caenorhabditis elegans Metabolism.', Cell Syst 2(5), 312--322. 2. Yilmaz, L. S. & Walhout, A. J. M. (2016), 'A Caenorhabditis elegans Genome-Scale Metabolic Network Model.', Cell Syst 2(5), 297--311.

Kissoyan, K.A.B. et al. (2019) International Worm Meeting "Natural C. elegans Microbiota Protects against Infection via Production of a Cyclic Lipopeptide of the Viscosin Group."

Dierking, K., Kissoyan, K.A.B.

[International Worm Meeting, 2019]

C. elegans is associated in nature with a species-rich, distinct microbiota, which was characterized only recently [1]. Our understanding of C. elegans microbiota function is thus still in its infancy. Here, we identify natural C. elegans microbiota isolates of the Pseudomonas fluorescens subgroup that increase C. elegans resistance to pathogen infection. We show that different Pseudomonas isolates provide paramount protection from infection with the natural C. elegans pathogen Bacillus thuringiensis through distinct mechanisms [2] . The P. lurida isolates MYb11 and MYb12 (members of the P. fluorescens subgroup) protect C. elegans against B. thuringiensis infection by directly inhibiting growth of the pathogen both in vitro and in vivo. Using genomic and biochemical approaches, we demonstrate that MYb11 and MYb12 produce massetolide E, a cyclic lipopeptide biosurfactant of the viscosin group, which is active against pathogenic B. thuringiensis. In contrast to MYb11 and MYb12, P. fluorescens MYb115-mediated protection involves increased resistance without inhibition of pathogen growth and most likely depends on indirect, host-mediated mechanisms. We are currently investigating the molecular basis of P. fluorescens MYb115-mediated protection using a multi-omics approach to identify C. elegans candidate genes involved in microbiota-mediated protection. Moreover, we are further exploring the antagonistic interactions between C. elegans microbiota and pathogens. This work provides new insight into the functional significance of the C. elegans natural microbiota and expands our knowledge of immune-protective mechanisms. 1. Zhang, F., Berg, M., Dierking, K., Felix, M.A., Shapira, M., Samuel, B.S., and Schulenburg, H. (2017). Caenorhabditis elegans as a model for microbiome research. Front. Microbiol. 8:485. 2. Kissoyan, K.A.B., Drechsler, M., Stange, E.-L., Zimmermann, J., Kaleta, C., Bode, H.B., and Dierking, K. (2019). Natural C. elegans Microbiota Protects against Infection via Production of a Cyclic Lipopeptide of the Viscosin Group. Curr. Biol. 29.

Christoph Janiesch et al. (2006) European Worm Meeting "Develoment Specific Collaboration between the E3 Ligases UFD-2 and CHN-1 Regulates the Myosin Assembly Chaperone UNC-45"

Johnny Kim, Christoph Janiesch, Thorsten Hoppe

[European Worm Meeting, 2006]

Christoph Janiesch, Johnny Kim and Thorsten Hoppe . In muscle, selective protein degradation by the ubiquitin-proteasome system is required to mediate the destruction of the sarcomeric structure, to regulate the maintenance and remodelling of the sarcomere and to ensure the development of muscle, the major component of the myofibrillar apparatus. The myosin chaperone UNC-45 plays a crucial role in the assembly of myosin into thick filaments and our recent work revealed that UNC-45 protein levels are subject to stringent regulation by two specific E3 ubiquitin ligases, CHN-1 and UFD-2. These two E3 enzymes form a novel E4 complex responsible for the multiubiquitylation of UNC-45, earmarking it for terminal protein degradation by the 26S proteasome.. . We have further investigated the roles of CHN-1 and UFD-2 during the process of UNC-45 dependent myosin assembly in C. elegans. Our current investigations revealed that movement defects of unc-45 thermosensitive (ts) mutants are suppressed in animals lacking either CHN-1 or UFD-2 and perturbation of UNC-45 protein level regulation by CHN-1 and UFD-2 results in severe movement defects specifically at the transition from L4 to young adult larval stage. This suggested that UNC-45 might be regulated in vivo by muscle specific co-expression of both E3 ubiquitin ligases in a developmentally regulated manner. We could support this notion by showing that UNC-45 protein levels increase during larval development and is highest at the L4 larval stage during the exponential growth phase of muscle thick filaments and degraded thereafter. Northern blot analysis identified an up-regulation of both chn-1 and ufd-2 transcripts specifically at the young adult larval stage, after body wall muscle development has occurred. In addition, our data suggest that unc-45, chn-1 and ufd-2 are not involved in myosin disassembly, suggesting their role to be specific for the developmental process but not for the degradation or reorganization of the sarcomere.

Johnny Kim et al. (2006) European Worm Meeting "Developmental Regulation of UNC-45 Mediated Myosin Assembly"

Thorsten Hoppe, Johnny Kim, Christoph Janiesch

[European Worm Meeting, 2006]

Johnny Kim, Christoph Janiesch and Thorsten Hoppe . In muscle, selective protein degradation by the ubiquitin-proteasome system is required to mediate the destruction of the sarcomeric structure, to regulate the maintenance and remodelling of the sarcomere and to ensure the development of muscle, the major component of the myofibrillar apparatus. The myosin chaperone UNC-45 plays a crucial role in the assembly of myosin into thick filaments and our recent work revealed that UNC-45 protein levels are subject to stringent regulation by two specific E3 ubiquitin ligases, CHN-1 and UFD-2. These two E3 enzymes form a novel E4 complex responsible for the multiubiquitylation of UNC-45, earmarking it for terminal protein degradation by the 26S proteasome.. We have further investigated the roles of CHN-1 and UFD-2 during the process of UNC-45 dependent myosin assembly in C. elegans. Our current investigations revealed that movement defects of unc-45 thermosensitive (ts) mutants are suppressed in animals lacking either CHN-1 or UFD-2 and perturbation of UNC-45 protein level regulation by CHN-1 and UFD-2 results in severe movement defects specifically at the transition from L4 to young adult larval stage. This suggested that UNC-45 might be regulated in vivo by muscle specific co-expression of both E3 ubiquitin ligases in a developmentally regulated manner. We could support this notion by showing that UNC-45 protein levels increase during larval development and is highest at the L4 larval stage during the exponential growth phase of muscle thick filaments and degraded thereafter. Northern blot analysis identified an up-regulation of both chn-1 and ufd-2 transcripts specifically at the young adult larval stage, after body wall muscle development has occurred. In addition, our data suggest that unc-45, chn-1 and ufd-2 are not involved in myosin disassembly, suggesting their role to be specific for the developmental process but not for the degradation or reorganization of the sarcomere. Importantly, both genetic and biochemical interactions of UNC-45, UFD-2 and CHN-1 are found from worm to man, indicating a conserved function of these components in muscle development across evolution.