Martin, Jack et al. (2021) International Worm Meeting "The kynurenine pathway is a major modulator of E. faecalis infection in C. elegans"

Whittingham-Dowd, Jayde, Cetnar, Kalina, Rezwana, Ruhi, Norvaisas, Povilas, Kosztelnik, Monika, Parry, Jackie, Au, Catherine, Martin, Jack, Cabreiro, Filipe, Zarate Potes, Alejandra, Urbaniak, Mick, Gems, David, Fathallah, Nadin, Hardgrave, Alex, Benedetto, Alexandre

[International Worm Meeting, 2021]

Key words Kynurenine pathway, E. faecalis, gut infection, microbiota, lysosome-related organelles autofluorescence. Abstract The kynurenine pathway (KP), main catabolic route for the essential amino-acid tryptophan, is well-known for its immunomodulatory role in mammals. While investigating death fluorescence in C. elegans, anthranilic acid (AA)-loaded lysosome-related organelles (LROs) were previously found responsible for the blue auto-fluorescence seen in the worm gut (Coburn et al. PLOS Biol. 2013). Given the bacteriostatic potential of AA and other kynurenine pathway compounds, we hypothesised that LROs and the KP play a key role in C. elegans gut microbial control. To test this idea, we exposed C. elegans to a worm-pathogenic strain of E. faecalis (OG1RF) and observed changes in gut morphology and autofluorescence dynamics upon infection. Transcriptomics and targeted metabolomics analyses further showed that KP activity is modulated upon E. faecalis infection. Using a combination of KP mutants from the Nollen lab (Van Der Goot et al. PNAS 2012), we observed that inhibition of various KP enzymes differentially affect C. elegans resistance to E. faecalis infection. E. faecalis growth on KP mutant worm extracts confirmed that resistant mutants produce bacteriostatic compounds, which we measured by HPLC. This was verified by the delayed or reduced gut colonisation of OG1RF-GFP (gifted by D. Garsin), and the ability for some mutants to thrive on OG1RF loans. We are currently investigating a broader role for the KP in C. elegans gut microbiota control, notably using newly generated CeMBio fluorescent strains.

Takeuchi T et al. (2011) Biol Pharm Bull "-galactosidases from jack bean and Streptococcus have different cleaving abilities towards fucose-containing sugars."

Kasai K, Natsuka S, Takeuchi T, Arata Y, Sugiura K, Nishiyama K, Takahashi H, Natsugari H

[Biol Pharm Bull, 2011]

We examined the sugar-cleaving abilities of -galactosidases from jack bean and Streptococcus towards sugars containing fucose residues, and found that jack bean -galactosidase has an ability to cleave the 1-3 linkage between galactose (Gal) and fucose (Fuc) residues, but not 1-4 linkage. On the other hand, streptococcal -galactosidase was found to cleave the linkage in both Gal1-4Fuc and Gal1-3Fuc disaccharide units. Such a difference in sugar-cleaving abilities between these 2 -galactosidases will be useful for structural analysis of glycans, especially those from species belonging to Protostomia, such as Caenorhabditis elegans.

Bowerman BA et al. (1994) Worm Breeder's Gazette "The Maternal Gene skn-4 and the Specification of Ventral Blastomere Fates in the Early C. elegans Embryo"

Bowerman BA, Shelton CA, Thorpe CJ, Martin PR

[Worm Breeder's Gazette, 1994]

THE MATERNAL GENE SKN-4 AND THE SPECIFICATION OF VENTRAL BLASTOMERE FATES IN THE EARLY C. ELEGANS EMBRYO Bruce Bowerman, Paula R. Martin, Christopher J. Thorpe, and Christopher A. Shelton. The Institute of Molecular Biology, University of Oregon, Eugene, OR 97403.

Zimmer M (2009) Chem Soc Rev "GFP: from jellyfish to the Nobel prize and beyond."

Zimmer M

[Chem Soc Rev, 2009]

On December 10, 2008 Osamu Shimomura, Martin Chalfie and Roger Tsien were awarded the Nobel Prize in Chemistry for "the discovery and development of the green fluorescent protein, GFP". The path taken by this jellyfish protein to become one of the most useful tools in modern science and medicine is described. Osamu Shimomura painstakingly isolated GFP from hundreds of thousands of jellyfish, characterized the chromophore and elucidated the mechanism of Aequorean bioluminescence. Martin Chalfie expressed the protein in E. coli and C. elegans, and Roger Tsien developed a palette of fluorescent proteins that could be used in a myriad of applications.

Hall DH et al. (1994) Worm Breeder's Gazette "Cytology of Degenerin-Induced Cell Death in the PVM Neuron"

Driscoll MA, Chalfie M, Gu GQ, Hall DH, Gong L

[Worm Breeder's Gazette, 1994]

Cytology of degenerin-induced cell death in the PVM neuron David H. Hall, Guoqiang Gu+, Lei Gong#, Monica Driscoll#, and Martin Chalfie+, * Dept. Neuroscience, Albert Einstein College of Medicine, Bronx, N.Y. 10461 + Dept. Biological Sciences, Columbia University, New York, N.Y. 10027 # Dept. Molecular Biology and Biochemistry, Rutgers University, Piscataway, N.J. 08855

Ray C et al. (1990) Worm Breeder's Gazette "CORRECTION Isolation of a C. elegans Cysteine Protease Gene"

McKerrow JH, Ray C

[Worm Breeder's Gazette, 1990]

In the previous issue of the WBG (Vol. 11, #3, page 20) we reported the isolation of a cysteine protease clone from a mixed-stage C. elegans cDNA library. This library was originally obtained from Chris Martin and not from Cynthia Kenyon as was reported in the article. Our apologies for any misunderstanding caused by this oversight.

Zarate Potes, Alejandra et al. (2021) International Worm Meeting "Solving host-microbe interactions and gut dysbiosis"

Benedetto, Alexandre, Zarate Potes, Alejandra, Ali, Irtiqa, Martin, Jack

[International Worm Meeting, 2021]

Gut host-microbe interactions critically impact C. elegans health and ageing. Multiple pathways and interactions have already been characterised in probiotic and dysbiotic contexts. However, we are still to unravel the complex host-microbe core or pathogen-specific genetic networks that orchestrate these interactions. We recently observed that the kynurenine pathway, main catabolic route for tryptophan, is majorly involved in regulating host-microbe interactions in worms, while insulin/IGF1 signalling (IIS) is a major upstream determinant of C. elegans gut biology and reproduction, immunity and longevity trade-offs. To solve part of this picture, we are now performing microbial and host transcriptomics, proteomics, and Tryptophan metabolite measurements in daf-2(e1370) and wild-type worms exposed to E. faecalis or P. aeruginosa for up to 12h, combined with host genome-wide RNAi infection screens relying on Label-Free Survival Assays (Benedetto et al. Aging 2019). In this poster we present the result of our bioinformatic investigations, main experimental pipelines and project strategy.

Ali, Irtiqa et al. (2021) International Worm Meeting "Investigating the basis of severe stress resistance in ageing"

Ackerley, Jacob, Benedetto, Alexandre, Ali, Irtiqa, Zarate-Potes, Alejandra, Martin, Jack

[International Worm Meeting, 2021]

Key words Severe stress resistance, ageing, CeMbio, IIS, oxidative, heat. Abstract Stress resistance and longevity are robustly correlated across heat stress paradigms but not oxidative stress paradigms. In particular, severe stress resistance in ageing C. elegans markedly differs between exposure to oxidative and heat stress that kill wild type worms within a couple of hours (Benedetto et al. Aging Cell 2019). Severe stress resistance may engage multiple mechanisms such as basal stress handling vs adaptive stress response pathways, different stress transduction pathways, the ability to execute the organismal death program (Galimov et al. 2019), or protection/sensitisation conferred by the gut microbiota. To disentangle these and evaluate their relative roles in ageing worms, we are screening C. elegans mutant in combination with strains from CeMBio collection for resistance to 7% tert-butyl hydroperoxide (t-BHP) and 42oC heat-shock as severe oxidative stress and thermal stress paradigms, respectively, before contrasting these results with aged animals. We are performing these screens on young adults at first, before moving into aged worms. So far, we have found that the capacity to execute the organismal death pathway does not significantly impact severe stress resistance in young adults fed an OP50 diet. However, young adults fed on 48 different bacterial isolates displayed varying levels of severe heat and oxidative stress resistance, some of which were insulin/IGF1-signalling (IIS) pathway-dependent. We are currently investigating the basis of these differences. This poster will provide an overview of the methods used, current results and planned future work.

Goto S (1999) Journal of Gerontology "Commentary on "Protein carbonyl accumulation in aging Dauer formation-defective (daf) mutants of Caenorhabditis elegans"."

Goto S

[Journal of Gerontology, 1999]

In recent years, oxidative damage to macromolecules has gained popularity as the basis of the molecular mechanism of aging. Martin proposes oxidative damage to macromolecules as one of the major public mechanisms of aging. Interest in modifications of protein by reactive oxygen species in aging was apparently introduced by Stadtman. Although various types of oxidative modifications can occur in proteins, carbonyl residues believed to be generated by metal catalyzed reaction or otherwise introduced by lysine, arginine and/or proline residues in vivo are often used as a marker of direct or

Hampoelz B et al. (2004) Cell "Heterotrimeric G proteins: new tricks for an old dog."

Knoblich JA, Hampoelz B

[Cell, 2004]

Heterotrimeric G proteins are well known for their function in signal transduction downstream of seven transmembrane receptors. More recently, however, genetic analysis in C. elegans and in Drosophila has revealed a second, essential function of these molecules in positioning the mitotic spindle and attaching microtubules to the cell cortex. Five new publications in Cell (Afshar et al., 2004; Du and Macara, 2004 [this issue of Cell]; Hess et al., 2004), Developmental Cell (Martin-McCaffrey et al., 2004), and Current Biology (Couwenbergs et al., 2004) show that this function is conserved in vertebrates and-like the classical pathway- involves cycling of G proteins between GDP and GTP bound conformations.