Yeast-based screening of natural product extracts results in the identification of prion inhibitors from a marine sponge

ABSTRACT

One of the major medical challenges of the twenty-first century is the treatment of incurable and fatal neurodegenerative disorders caused by misfolded prion proteins. Since the discovery of these diseases a number of studies have been conducted to identify small molecules for their treatment, however to date no curative treatment is available. These studies can be highly expensive and time consuming, but more recent experimental approaches indicate a significant application for yeast prions in these studies. We therefore used yeast prions to optimize previous high-throughput methods for the cheaper, easier and more rapid screening of natural extracts. Through this approach we aimed to identify natural yeast-prion inhibitors that could be useful in the development of novel treatment strategies for neurodegenerative disorders. We screened 500 marine invertebrate extracts from temperate waters in Australia allowing the identification of yeast-prion inhibiting extracts. Through the bioassay-driven chemical investigation of an active Suberites sponge extract, a group of bromotyrosine derivatives were identified as potent yeast-prion inhibitors. This study outlines the importance of natural products and yeast prions as a first-stage screen for the identification of new chemically diverse and bioactive compounds.     

Sterol composition of the marine sponge Crambe crambe

The sterol content of the marine sponge Crambe crambe has been determined. The major components of the mixture are cholest-7-en3β-ol, 24-methylcholesta-7,22-dien-3β-ol and cholesta-7,22-dien-3β-ol. Significative quantities of the rare 4α-methyl-5α-cholest-8-en3β-ol are also present.     

The taxonomic and ecological status of the environmentally restricted spongillid species of North America. I. Spongilla sponginosa Penney 1957

The taxonomic validity, present distribution, and specific threats to the existence of the freshwater sponge,Spongilla sponginosa Penney were investigated. This species, reported only from the type locality, Week's Pond, Sumter County, South Carolina, has apparently been extirpated due to highly acidic pH levels in the pond water. Examination of holotype materials indicate some question of the validity of S. sponginosa as a distinct species.     

Plectroninia celtica n. sp. (Calcarea, Minchinellidae), a new species of “pharetronid” sponge from bathyal depths in the northern Porcupine Seabight, NE Atlantic

Recent pharetronid sponges were regarded as relict species in tropical and subtropical waters, inhabiting cryptic habitats on coral reefs and in caves. More recent findings of a new species of the genus Plectroninia off northern Norway, with an inner fused skeleton have changed that view. Recent investigations on the sponge fauna of the “Propeller Mound”, northern Porcupine Seabight, focusing on sponges growing on the azooxanthellate cold-water coral Lophelia pertusa (Linné 1758) and Madrepora oculata Linné 1758, established the presence of a species of Plectroninia new to science. Its status as a common species within this deep-water coral habitat and the general status of the genus Plectroninia are discussed.     

A modern approach to demineralization of spicules in glass sponges (Porifera: Hexactinellida) for the purpose of extraction and examination of the protein matrix

Glass sponges of the class Hexactinellida are a group of the most ancient multicellular animals, whose fossil remnants from the early Proterozoic have been registered. In order to demineralize the skeletal structures of the glass sponge Hyalonema sieboldi, we have used for the first time a strategy of slow leaching of the silicon-bearing component, based on the usage of alkaline solutions of sodium hydroxide, sodium dodecyl sulfate, and an anionic biosurfactant of a rhamnolipid nature. The obtained data unequivocally corroborate the presence of a fibrillar protein matrix functioning as a basis for silicon biomineralization in the basal spicules of H. sieboldi. Also, it has been found for the first time that the protein matrix is constructed of a collagenous protein. The technical approach proposed here might appear important for the study of the structural organization of skeletons in other silicon-bearing animals and, in an applied aspect, to work out new biomaterials for implantology and biocomposites, in order to use the latter as bioactive additives.     

Species-Specific Association of the Cell-Aggregation Molecule Mediates Recognition in Marine Sponges

Reaggrcgation of dissociated cells of marine sponges, resulting in reformation of functional sponges, is a calcium-dependent process mediated by large, proteoglycan-like molecules termed aggregation factors (AF). During aggregation, species-specific sorting of cells is often observed. We purified and characterized AFs from three different sponge species and investigated their role in species-specific aggregation using novel approaches. The calcium-dependent association between purified AFs is species-specific in most combinations, as was shown in overlay assays and bead-sorting assays with AFs immobilized onto colored beads. Species-specific interactions of living cells and AF-beads resulted in incorporation of only homospecific AF-beads into reforming cell aggregates. Sequences from peptides obtained from the AF core proteins could all be aligned to the sequence of one species, the Microciona prolifera AFp3 core protein. In contrast to this similarity, major species-specific differences were seen in carbohydrate composition and in the response of AFs to specific carbohydrate-recognizing antibodies. In summary, our data point to a prominent role for the calcium-dependent association of AFs in recognition processes during aggregation. As this association of AFs occurs via carbohydrate -carbohydrate interactions, we speculate that the specificity of those interactions may be fundamental to recognition mechanisms required for regeneration of individuals from dissociated cells and for rejection of foreign material by sponge individuals.     

Discordance between morphological and molecular species boundaries among Caribbean species of the reef sponge Callyspongia

Sponges are among the most species‐rich and ecologically important taxa on coral reefs, yet documenting their diversity is difficult due to the simplicity and plasticity of their morphological characters. Genetic attempts to identify species are hampered by the slow rate of mitochondrial sequence evolution characteristic of sponges and some other basal metazoans. Here we determine species boundaries of the Caribbean coral reef sponge genus Callyspongia using a multilocus, model‐based approach. Based on sequence data from one mitochondrial (COI), one ribosomal (28S), and two single‐copy nuclear protein‐coding genes, we found evolutionarily distinct lineages were not concordant with current species designations in Callyspongia. While C. fallax, C. tenerrima, and C. plicifera were reciprocally monophyletic, four taxa with different morphologies (C. armigera, C. longissima, C. eschrichtii, and C. vaginalis) formed a monophyletic group and genetic distances among these taxa overlapped distances within them. A model‐based method of species delimitation supported collapsing these four into a single evolutionary lineage. Variation in spicule size among these four taxa was partitioned geographically, not by current species designations, indicating that in Callyspongia, these key taxonomic characters are poor indicators of genetic differentiation. Taken together, our results suggest a complex relationship between morphology and species boundaries in sponges.     

Reproductive cycles of the sympatric excavating sponges Cliona celata and Cliona viridis in the Mediterranean Sea

Abstract. Clionaids are excavating sponges, which live in and grow into calcareous substrates. We studied the sexual reproductive cycles of two clionaid sponges coexisting in a Mediterranean coastal basin (Porto Cesareo, Italy), Cliona viridis and Cliona celata, by analyzing monthly tissue samples of ten specimens of each species collected over a 2‐year period. From May to June of the second study year, supplementary samples were taken weekly. Up to 90% of the specimens of C. viridis and 70% of those of C. celata sampled were reproductive during the study. In both species, but particularly in C. viridis, reproductive investment, measured as the percentage of sponge tissue occupied by gametes, was high. Oocytes were present almost year‐round in both species, except for a 1–4‐month period after zygote release. In contrast, spermatogenesis occurred most frequently in May in both species, when (May–June) oocytes reached their greatest diameters. Cliona viridis and C. celata are hermaphrodites, with oocytes and spermatic cysts coexisting in 10% of the studied individuals in the first year of the study, and in 30% during the second. No developing embryos or larvae were incubated in the sponge tissues, and fertilization was not observed. Temperature may play a role in triggering some important phases of the reproduction of these Cliona, such as oocyte maturation and spermatogenesis, which occurred when water temperature increased from 17°C to 25°C between May and June.     

Bio-sintering processes in hexactinellid sponges: Fusion of bio-silica in giant basal spicules from Monorhaphis chuni

The two sponge classes, Hexactinellida and Demospongiae, comprise a skeleton that is composed of siliceous skeletal elements (spicules). Spicule growth proceeds by appositional layering of lamellae that consist of silica nanoparticles, which are synthesized via the sponge-specific enzyme silicatein. While in demosponges during maturation the lamellae consolidate to a solid rod, the lamellar organization of hexactinellid spicules largely persists. However, the innermost lamellae, near the spicule core, can also fuse to a solid axial cylinder. Similar to the fusion of siliceous nanoparticles and lamella, in several hexactinellid species individual spicules unify during sintering-like processes. Here, we study the different stages of a process that we termed bio-sintering, within the giant basal spicule (GBS) of Monorhaphis chuni. During this study, a major GBS protein component (27 kDa) was isolated and analyzed by MALDI-TOF-MS. The sequences were used to isolate and clone the encoding cDNA via degenerate primer PCR. Bioinformatic analyses revealed a significant sequence homology to silicatein. In addition, the native GBS protein was able to mediate bio-silica synthesis in vitro. We conclude that the syntheses of bio-silica in M. chuni, and the subsequent fusion of nanoparticles to lamellae, and finally to spicules, are enzymatically-driven by a silicatein-like protein. In addition, evidence is now presented that in hexactinellids those fusions involve sintering-like processes.     

The Sula Reef Complex, Norwegian shelf

Summary Cool-water carbonates in the aphotic zone of deep shelf and continental margin settings in the Northeast Atlantic are produced by the deep-water coral reefs withLophelia pertusa as the major framework builder. Through a compilation of side scan sonar, airgun and manned submersible surveys from several cruises to the mid-Norwegian Sula Reef Complex (SRC), the facies pattern and zonation of one of the largest deep-water reefs in the Northeast Atlantic is described in relation to the overall seabed topography. The late glacial to early postglacial iceberg scour on the crest and shoulder of the Sula Ridge provides settling ground for the scleractinian corals already in the early Holocene. Since then coral growth continues until today but was supposed to be disturbed by an environmental hazard, the so-called second Storegga event. The distinct distribution pattern of individualLophelia reefs on the Sula Ridge has stimulated a discussion on intrinsic environmental controls such as the bentho-pelagic coupling and the alternative hydrocarbon-based nutrition hypothesis.