ABSTRACT: LC-MS runs of sponge extracts from Theonella swinhoei sponges (white chemotype) collected in Israel and Japan with a focus on theonellamide production
Project description:In the seabed, chemical defences mediate inter- and intraspecific interactions and may determine organisms’ success, shaping the diversity and function of benthic communities. Sponges represent a prominent example of chemically-defended marine organisms with great ecological success. The ecological factors controlling the production of their defensive compounds and the evolutionary forces that select for these defences remain little understood. Each sponge species produces a specific and diverse chemical arsenal with fish-deterrent, antifouling and antimicrobial properties. However, some small animals (mesograzers), mainly sea slugs, have specialized in living and feeding on sponges. Feeding on chemically-defended organisms provides a strategy to avoid predators, albeit the poor nutritional value of sponges. In order to investigate the mechanisms that control sponge chemical defence, with particular focus on the response to specialist grazers, we investigated the interaction between the sponge Aplysina aerophoba and the sea slug Tylodina perversa. Here we performed controlled experiments and collected sponge samples at different time points (3h, 1d and 6d after treatment). To further elucidate if the sponge response is specific to grazing by T. perversa, we also included a treatment in which sponges were mechanically damaged with a scalpel. We compared gene expression between treatments based on RNA-Seq data.
Project description:Tropical lagoon-inhabiting organisms live in highly irradiated ecosystems and are particularly susceptible to thermal stress resulting from climate change. However, despite living close to their thermal maxima, stress response mechanisms found in these organisms are poorly understood. We used a novel physiological-proteomic approach for sponges to describe the stress response mechanisms of the lagoon-inhabiting sponge Amphimedon navalis, when exposed to elevated seawater temperatures of +2 oC and +4 oC relative to a 26 oC ambient temperature for four weeks. After four weeks of thermal exposure, the buoyant weight of the sponge experienced a significant decline, while its pumping rates and oxygen consumption rates significantly increased. Proteome dynamics revealed 50 differentially abundant proteins in sponges exposed to elevated temperature, suggesting that shifts in the sponge proteome were potential drivers of physiological dysfunction. Thermal stress promoted an increase in detoxification proteins, such as catalase and glutathione-S-transferase, suggesting that an excess of reactive oxygen species in sponge cells were likely responsible for the significant increase in oxygen consumption. Elevated temperature also disrupted cellular growth and cell proliferation, promoting the loss of sponge biomass, and the high abundance of multiple alpha-tubulin chain proteins also indicated an increase in cytoskeletal activities within sponge cells, which may have induced the increase in sponge pumping rate. Our results show that sustained thermal exposure in susceptible lagoonal sponges may induce significant disruption of cellular homeostasis leading to physiological dysfunction, and that a combined physiological-proteomic approach may provide new insights into physiological functions and cellular processes occurring in sponges.
Project description:Demosponge Cinachyrella cf cavernosa is an inter-tidal sponge. It is found in competition with soft coral Zoanthus sansibaricus and macroalgae Dictyota ciliatum. The effect of these two spatial competitors on the gene expression profile of the sponge is checked. Sponges are collected from three distinct situations, 1. sponge without competitors, 2. sponge in competition with algae, and 3. sponge in competition with soft coral. Each group has three biological replicates.
Project description:UPLC-MS/MS or nanoLC-MS/MS data acquired from whole sponge extracts from the marine sponge Theonella swinhoei (yellow chemotype; Hachijo Jima, Japan) or extracts from two Entotheonella enriched cell fractions obtained by differential centrifugation. More than 40 bioactive polyketides and modified peptides that belong to seven structural classes are known to be produced by this sponge. Single-cell genomics and chemical analysis have revealed Entotheonella as the producer of most of the known chemistry from this sponge.
Project description:This work reveals the deeply conserved gene repertoire of animal stem cells, from sponges to mammals. mRNA profiles totipotent stem cells (archeocytes), choanocytes, other differentiated cell types in a freshwater sponge and were mapped on the reference transcriptome generated in the same study
Project description:The interaction of animals with microbes relies on the specific recognition of microbial-derived molecules by receptors of the immune system. Sponges (phylum Porifera), as sister group of the Eumetazoa, provide insights into conserved mechanisms for animal-microbe crosstalk, but empirical data is limited. Here we aimed to characterize the immune response of sponges upon microbial stimuli by RNA-Seq. Two sponges species from the Mediterranean Sea, Aplysina aerophoba and Dysidea avara, were challenged with microbial-associated molecular patterns (lipopolysaccharide and peptidoglycan) or sterile artificial seawater (control) in aquarium experiments. Sponge tissue samples were collected 1h, 3h, and 5h after treatment. The response of the sponges to the treatments was assessed by differential gene expression analysis of RNA-Seq data. For each species, we compared the transcriptomic profiles of the samples in MAMP treatment to control within each time point.
Project description:This dataset contains several sponge-derived bacteria, along with data from the sponges to enable the assignation of the biosynthetic source of detected metabolite features. Media controls and solvent blanks are also included. Please see metadata for the bacterial source sponge.
Project description:Ephydatia muelleri is a cosmopolitan freshwater demosponge, with potential to become a model system. We have participated in a large collaborative project to sequence the genome (PRJNA579531), methylome, transcriptome for this species, aiming to better understand the biology of this sponge species. In terms of DNA methylation, it presents relatively low methylation levels compared to the methylomes of other sponges (A. queenslandica and S. ciliatum), suggesting quite a lot of varation within the sponge phylum.
Project description:Marine sponges represent one of the few eukaryotic groups that ubiquitously harbor symbiotic members of the Thaumarchaeota, which are important chemoautotrophic ammonia-oxidizers in many environments. However in most studies, direct demonstration of ammonia-oxidation by these archaea within sponges is lacking, and little is known about sponge-specific adaptations of archaeal ammonia oxidizers (AOA). In this study, we characterized the thaumarchaeal symbiont of the marine sponge Ianthella basta using metaproteogenomics, fluorescence in situ hybridization, qPCR and direct isotope-based functional assays. We demonstrate that the I. basta symbiont is not closely related to other genomically sequenced sponge AOA and is a member of a new genus. “Candidatus Nitrosospongia bastadiensis” is an abundant symbiont that is solely responsible for nitrite formation from ammonia in I. basta that surprisingly does not harbor nitrite-oxidizing microbes. Consistently, Ca N. bastadiensis encodes and expresses the genetic repertoire required for chemolithoautotrophic ammonia oxidation. Furthermore, we show that this AOA is equipped with an expanded set of extracellular subtilisin-like proteases, a metalloprotease unique among archaea, as well as a putative branched-chain amino acid ABC transporter. This repertoire is strongly indicative of a mixotrophic lifestyle and is (with slight variations) also found in other sponge-associated, but not in free-living AOA. We predict that this feature as well as an expanded and unique set of secreted serpins (protease inhibitors), a unique array of eukaryotic-like proteins, and a DNA-phosporothioation system likely involved in defense against foreign DNA, represent important adaptations of AOA to life within these ancient filter-feeding animals.
Project description:MicroRNA (miRNA) sponges containing miRNA complementary binding sites constitute a potentially useful strategy for miRNA-inhibition therapeutics in cancer patients. Recently, naturally occurring circular RNAs (circRNAs) have been revealed to function as efficient microRNA sponges. We hypothesized that synthetic circRNA sponges targeting oncomiRs could be constructed and used to achieve potentially therapeutic microRNA loss of function. In this study, linear RNA molecules containing five miR-21 binding sites were transcribed in vitro. After dephosphorylation by calf intestinal phosphatase and phosphorylation by T4 polynucleotide kinase, circRNA sponges were circularized using 5’-3’ end ligation by T4 RNA ligase 1. Synthetic circular sponge stability was assayed in the presence of RNase R or fetal bovine serum. Luciferase reporter and cell proliferation assays were performed to assess competitive inhibition of miR-21 activity by circRNA sponges in NCI-N87 gastric cancer cells. Tandem Mass Tag (TMT) labeling proteomics analysis and Western blotting were performed to delineate effects of circRNA sponges on miR-21 downstream targeted proteins. Our experiments revealed that artificial circRNA sponges can be synthesized using enzymatic ligation. These synthetic circRNA sponges are more resistant than their linear RNA counterparts to nuclease degradation in vitro. They effectively suppress the activity of miR-21 on its downstream protein targets, including the important cancer protein DAXX. Finally, they also inhibit gastric cancer cell proliferation. Our results suggest that synthetic circRNA sponges represent a rapid, effective, convenient strategy to achieve loss of miRNA function in vitro, with potential future therapeutic application in vivo.