Stone AL et al. (1999) J Protein Chem "Structure-function analysis of the ADAM family of disintegrin-like and metalloproteinase-containing proteins (review)."

Sang QX, Stone AL, Kroeger M

[J Protein Chem, 1999]

The ADAMs belong to a disintegrin-like and metalloproteinase-containing protein family that are zinc-dependent metalloproteinases. These proteins share all or some of the following domain structure: a signal peptide, a propeptide, a metalloproteinase, a disintegrin, a cysteine-rich, and an epidermal growth factor (EGF)-like domains, a transmembrane region, and a cytoplasmic tail. ADAMs are widely distributed in many organs, tissues, and cells, such as brain, testis, epididymis, ovary, breast, placenta, liver, heart, lung, bone, and muscle. These proteins are capable of four potential functions: proteolysis, adhesion, fusion, and intracellular signaling. Because the number of ADAM genes has grown rapidly and the biological functions of most members are unclear, this review analyzes the protein structures and functions, their activation and processing, their known and potential activities, and their evolutionary relationships. A sequence alignment of human ADAMs is compiled and their homology and physical data are calculated. The conceivable functions of ADAMs in reproduction, development, and diseases are also discussed.

Coates D et al. (2000) European Worm Meeting "The ADAMs family of C. elegans: what will the new members do?"

Coates D, Daulby J, Hooper NM, Hope IA

[European Worm Meeting, 2000]

ADAMs are a family of integral membrane glycoproteins containing a disintegrin and a metalloprotease domain. The physiological roles of the majority of ADAMs have yet to be elucidated, however some mammalian ADAMs are known to be involved in many diverse processes including sperm migration, sperm-egg binding and fusion, myoblast fusion, the processing of adhesion molecules, cytokines, cytokine receptors and extracellular protein domains, and in neural development. In C. elegans ADAMs are thought to be involved in cell fusion events in sperm and in epithelial cells (ADM-1), in early embryonic development (ADM-2) and in vulval development (SUP-17). In addition to these membrane anchored proteins C. elegans also has soluble ADAM-like proteases, for example GON-1, which plays an essential role in gonadal morphogenesis. We have identified four novel ADAM-like sequences in C. eleganswhich encode a TNFa converting enzyme (TACE) homologue and three soluble ADAM-like proteinases, and our aim is to determine the precise functions of these proteins. Initially we will examine their cellular location with reporter gene constructs and assess the importance of these proteinases by generating gene knockout mutants. We are also interested in finding out what the substrates of these ADAMs are and will biochemically characterise the recombinant proteins. The data we will attain will be related to the ADAM homologues found in humans.

Crawley M et al. (1998) Worm Breeder's Gazette "Correction"

Adams D, Crawley M

[Worm Breeder's Gazette, 1998]

Correction: "The abstract entitled Control of Mitochondrial Morphology published in WBG Vol. 15 No. 1 contained an error. Worms were treated with 3 mM (not 100 mM) chloramphenicol to induce changes in mitochondrial morphology. Thanks to A.L. for spotting the error. M. Crawley & D. Adams

Adir V et al. (1997) International C. elegans Meeting "ADM-2 A NEW MEMBER OF THE ADAM FAMILY WITH AN ACTIVE METALLOPROTEASE DOMAIN"

Podbilewicz B, Adir V

[International C. elegans Meeting, 1997]

The ADAMs gene family encodes transmembrane proteins which contain both disintegrin and metalloprotease domains. Some ADAMs have a metalloprotease like domain but lack the active site, while others have a consensus active site sequence for a zinc dependent metalloprotease as in snake venoms. The ADAMs gene family includes fertilin a and b (involved in sperm-egg fusion), meltrin-a (required for myotube formation in mice), TACE (a TNF-a converting enzyme), and KUZ (the kuzbanian gene product essential for neural fate in Drosophila). Other members of this family may be involved in different cell-cell and cell-matrix interactions. ADM-1, was the first gene from this family found in C. elegans, and is expressed in cells that undergo cell fusion. However, ADM-1 lacks the consensus active site sequence for a zinc dependent metalloprotease. In order to identify new genes related to the ADAMs gene family in C. elegans, we used the TBLASTN algorithm and found the cDNA clone yk35e9 which encodes adm-2. Alignment of ADM-2 with the metalloprotease domains of other ADAMs, shows that the consensus active site is highly conserved. adm-2 was mapped to YACs Y50F10 and Y49B10 and to the cosmid C04A11. Consensus HEXXHXXGXXH- ADM-2 HELGHDFGNDHD fertilin a HELGHNLGIRHD (Blobel et al., Nature, 1992) meltrin a HELGHNFGMNHD (Yagami Hiromasa et al., Nature, 1995) TACE HELGHNFGAEHD (Black et al., Nature, 1997) KUZ HEIGHNFGSPHD (Rooke et al., Science, 1996) adamalysin HELGHNLGMEHD (Gomis-Ruth et al., EMBO J., 1993) ADM-1 QSIGHLLGLEHD (Podbilewicz, Mol. Biol. Cell, 1996) Alignment of the consensus active sites for zinc-dependent metalloproteases and ADM-1. To study the function of adm-2 and its effect on C. elegans development we inject adm-2 antisense RNA into wild type nematodes. We also perform rescue experiments to candidate mutants that mapped genetically to the genome location of adm-2.

Joanna Daulby et al. (2001) International C. elegans Meeting "Expression of C. elegans ADAM proteins in a mammalian system."

Nigel M Hooper, Joanna Daulby, David Coates, Ian A Hope

[International C. elegans Meeting, 2001]

ADAMs are integral membrane proteins which contain a d isintegrin a nd m etalloprotease domain, and have the potential to participate in proteolysis, adhesion, fusion and signaling events. So far thirty ADAM genes have been described from across the animal kingdom, but their precise functions and mechanisms remain largely unknown. A small number of mammalian ADAMs however, have been shown to be important in development, signal transduction, sperm and egg binding and muscle cell fusion. In addition, some C. elegans ADAMs have been investigated recently and these are thought to be involved in sperm and epithelial cell fusion events ( adm-1 ), in early embryonic development ( adm-2 ), and in vulval development ( sup-17 ). C. elegans also possesses homologues of ADAM-like genes known as ADAM-TS. Proteins encoded by this gene family are not membrane bound and have an additional thrombospondin-like motif, for example GON-1 and MIG-17. These proteins play essential roles in the morphogenesis of the C. elegans gonad. We are studying three other ADAMs genes in C. elegans : adm-4 (a tumour necrosis factor alpha converting enzyme or TACE homologue); and two soluble ADAM-TSs, ( C02B4.1 and T19D2.1 ). GFP reporter data indicates that adm-4 is highly expressed in most tissues in all post-embryonic stages. The expression pattern for C02B4.1 :: GFP however is more confined and is observed in body wall and pharyngeal muscle. We aim to complement the reporter data with RNAi/knockout mutagenesis work, but our main goal is to biochemically analyse the recombinant proteins in a mammalian expression system. Using this approach we hope to identify which proteins are cleaved by these ADAMs proteins, as well as providing information about their targeting and processing. Ultimately we hope to elucidate their physiological function and relate this information to mammalian ADAM proteins.

Lam, Arielle et al. (2015) International Worm Meeting "The identification of SYS-1/beta-catenin binding partners and transactivation domains in the Wnt/beta-catenin asymmetric pathway."

Molumby, Michael, Clemons, Amy, Phillips, Bryan, Lam, Arielle, Adams, Lori

[International Worm Meeting, 2015]

Wnt/beta-catenin signaling plays a crucial role in embryonic development. In worms, a similar pathway, Wnt/beta-catenin asymmetric (WbetaA) pathway, controls binary cell fate decisions resulting in asymmetric cell division (ACD). ACD produces two daughter cells with different signaling profiles, in which one daughter exhibits activation of WbetaA target genes while the other inactivates WbetaA signaling. SYS-1/beta-catenin, a transcriptional activator is a key protein in WbetaA pathway and is tightly regulated in both daughter cells. In the Wnt inactive cells, SYS-1 appears to be actively degraded while in the Wnt active daughter; SYS-1 is stabilized and transported into the nucleus to interact with POP-1/TCF transcription factors to activate Wnt dependent genes. Despite having poor sequence conservation to mammalian beta-catenin, SYS-1 exhibit close structural similarities and is shown to interact with various proteins within the WbetaA pathway in a similar manner to its mammalian counterpart. However, the transactivation domain(s), the binding sites of various SYS-1 regulators, as well as any SYS-1 post-translational modification all remain unknown. We therefore have undertaken a structure-function and mass spectrometry approach to identify SYS-1 transactivation domains as well as other unknown SYS-1 binding partners. Yeast-one-hybrid assay with SYS-1 truncations shows that the N-terminal domains as well as the first four armadillo repeats are able to activate transcription. When tested in worms, these two truncations linked to POP-1/TCF DNA binding domain are sufficient to induce the Wnt active fate in both daughters of an ACD. Preliminary mass spectrometry experiment done on worms overexpressing SYS-1 resulted in identifying several potential SYS-1 interactors. Further experiments will be done to verify the interactions between those proteins with SYS-1. These studies will help identify SYS-1 transactivation domains and novel SYS-1 interactors, which will aid in identifying novel mechanisms of Wnt target gene regulation.

Lam, Koonyee et al. (2013) International Worm Meeting "Structure-function of SYS-1/b-catenin, an effector of Wnt-directed asymmetric cell divisions."

Adams, Lori, Lam, Koonyee, Hutchinson, Jennifer, Molumby, Michael, Phillips, Bryan

[International Worm Meeting, 2013]

Wnt/b-catenin signaling is an important component in embryonic development as well as homeostasis of adult tissues. A related Wnt/b-catenin Asymmetry (WbA) pathway controls binary cell fate decisions during the many C. elegans asymmetric cell divisions (ACD). WbA requires the proper regulation and function of the transcriptional activator in this pathway, the b-catenin SYS-1. ACD produces two daughter cells with different SYS-1 expression profiles, with high SYS-1 expression in the Wnt responsive cell and low SYS-1 expression in the Wnt nonresponsive cell. Despite poor sequence conservation, the SYS-1 crystal structure shows close structural similarities with mammalian b-catenin, namely twelve armadillo repeats with a specific TCF binding domain. SYS-1 lacks an unfolded C-terminal domain (CTD) that is utilized by mammalian b-catenin for the recruitment of several transcriptional co-activators such as DPY-22/MED12 and CBP-1/CBP. Genetic analyses of these co-activators indicate they are required for proper ACD, suggesting they may be recruited to Wnt target genes by SYS-1 via a novel interaction. Likewise, SYS-1 lacks GSK3b and CK1a regulatory sites, which facilitate degradation of b-catenin. We therefore have undertaken a structure-function approach to identify SYS-1 transactivation and regulatory domains. Yeast one-hybrid assays with SYS-1 fragments show that the unfolded N-terminal domain (NTD) and the first four armadillo repeats, but not the CTD repeats, are able to separately activate transcription. In worms, these two activation domains, when linked to the POP-1/TCF DNA binding domain, are sufficient to induce the Wnt signaled fate in both daughters of an ACD. Consistent with a role in transcription, NTD, but not the CTD, fragments preferentially localize to C. elegans nuclei after ACD. Finally, no SYS-1 fragment has yet been identified that is sufficient to drive degradation in Wnt inactive cells, suggesting any modification sites are distinct from regulatory binding sites and that a targeting scaffold may be involved in SYS-1 degradation. These studies will help identify SYS-1 regulatory and transactivation domains, which will aid in identifying novel mechanisms of Wnt target gene regulation.

H Pamjav et al. (1999) International C. elegans Meeting "Phylogenetic analysis of entomopathogenic nematode / bacterium symbiotic complexes by using PhastSystem PAGE PCR-RFLP of rDNA spacer sequences"

D Triga, A Fodor, H Pamjav, Z Buzas, E Szallas

[International C. elegans Meeting, 1999]

Entomopathogenic nematode (EPN)species belonging to the Steinernema and Heterorhabditis genera are closely related to C. elegans (Blaxter et al., 1998). They form an intimate symbiotic complex with bacteria Xenorhabdus spp. and Photorhabdus spp., respectively, which are closely related to E. coli . The symbiosis is developmentally regulated: there is a direct cell-cell contact between the gut cell of the (infective) dauerlarva and the primary phase variant of the symbiotic bacteria. The symbiosis is taxon specific. Individual Xenorhabdus species used as symbionts by each Steinernema species and individual strains of Photorhabdus luminescens used as symbionts by each Heterorhabditis strains belonging to different species. The phylogenetic relations between species and strains have been analysed by RFLP of the internally transcribed spacer region of the respective rRNA operon. The PCR-amplified DNAs were digested by different restriction enzymes and the restriction patterns obtained by PhastSystem PAGE were compared. On the basis of the comparative analysis of the ITS1 - ITS2 patterns nematodes could be identified at species level. Xenorhabdus species could also be identified on the basis of the comparative analysis of the spacer region of the 16S - 23S rRNA operon. P. luminescens strains, belonging to different subclusters (Szallas et al., 1997) could be separated on the basis of the RFLP pattern of the spacer region of the 16S - 23S rRNA operon. Adams, B.J. (1998). Species conception and the evolutionary paradigm in modern nematology. J. nematol. 30, 1-21. Adams, B.J., Burnell, A.M. @ Powers,T.O. (1998). A phylogenetic analysis of Heterorhabditis based on ITS 1 DNA sequence data. J. nematol. 30, 22-39. Blaxter,R. et al.(1998).Molecular evolutionary framework for Phylum nematoda. Nature 392,71-75. Szallas E., Koch, H., Fodor,A.,Szentirmai, A., Nealson,K.H. Stackebrandt, E.(1997).Phylogenetic evidence for taxonomic heterogeneity of P.luminescens. IJSB 47, 402-407.

Ryan Viveiros et al. (2007) International Worm Meeting "An RNAi Screen to Identify Genes Involved in Myoblast Migration and Positioning in the Nematode Caenorhabditis elegans."

Harald Hutter, Ryan Viveiros, Donald Moerman

[International Worm Meeting, 2007]

In C. elegans myoblasts arise from several different founder cell lineages. These cells initially arrange themselves in two rows along the left and right lateral midlines and at ~290 min of development they migrate dorsally and ventrally to form the four muscle quadrants present upon hatching (Sulston et al,1983). As the myoblasts migrate they are still dividing, as are many other cells in their immediate environment. This means the cell-cell contact of cells during migration is dynamic and can vary from animal to animal (Schnabel et al, 1997). This situation creates an environment where the extracellular matrix (ECM) and cell surface contacts are in constant flux, which begs the questions as to how these cells navigate unerringly to their final destination. A number of ECM and cell surface components are known to affect cell positioning, migration and attachment, including laminin, basement membrane collagens, and the NCAM, cadherin and i ntegrin families (reviewed in Levi et al,1990, and Adams and Watt, 1993). The C. elegans homologues of these proteins are involved in these processes during later stages of development, but do not appear to have a role during myoblast migration. In an effort to identify the proteins involved in early myoblast migrations we first compiled a list of 948 genes encoding proteins predicted to be in, or to interact with the ECM. To date we have completed an initial screen of 776 genes using RNAi knockdowns and found that ~40% showed some phenotype, with ~8% of the positives showing some postembryonic muscle positioning, migration, or attachment aberration. We are currently retesting these preliminary positives and investigating whether the muscle phenotypes observed are tissue specific or represent more general defects. Adams, J. and Watt, F. (1993). "Regulation of development and differentiation by the extracellular matrix." Development 117(4): 1183-1198 Levi, G, et al. (1990). "Modes of cell migration in the vertebrate embryo." Int Rev Cytol(123): 201-52 Schnabel, R., et al. (1997). "Assessing normal embryogenesis in Caenorhabditis elegans using a 4D microscope: variability of development and regional specification." Dev Biol 184(2): 234-65 Sulston, J., et al. (1983). The embryonic cell lineage of the nematode Caenorhabditis elegans. Dev. Biol. 100(1): 64-119.

Noble, M. et al. (2017) International Worm Meeting "The Nucleosome Remodelling Factor subunit pyp-1 negatively regulates the heat shock response upon aging in Caenorhabditis elegans ."

Guisbert, E, Bowie, L, Deonarine, A., Westerheide, S, Lugano, D., Noble, M.

[International Worm Meeting, 2017]

Title: The Nucleosome Remodelling Factor subunit pyp-1 negatively regulates the heat shock response upon aging in Caenorhabditis elegans Mark Noble, Andrew Deonarine, Doreen Lugano, Lori-Ann Bowie and Sandy D. Westerheide The heat shock response is a conserved eukaryotic stress response known to regulate protein homeostasis (proteostasis) and it is regulated by the transcription factor hsf-1. Previous studies have shown tremendous effects of the HSR on life span, health span, and disease models of neurodegenerative diseases such as Alzheimer's, Parkinson and Huntington's disease. In C. elegans, proteostasis has been shown to decline with the onset of adulthood in parallel to the decline of the HSR. Therefore, understanding the molecular mechanisms and regulation of this response is vital in revealing its effects on various stresses. Recent studies have shown a link between chromatin remodelers and regulation of the heat shock response. Thus, utilizing RNAi genetic screen, we were able a uncover pyp-1, a pyrophosphatase and subunit of the Nucleosome-remodeling factor (NURF) which showed tremendous effects on the HSR. From a different screen, pyp-1 was also found to be a negative regulator of the heat shock response in C. elegans. We hypothesize that the age-dependent decrease in HSR induction may be due to repressive chromatin changes conferred by chromatin remodeling complexes. As a result, we identified pyp-1 as a novel negative regulator of the heat shock response and proteostasis upon aging in C.elegans.