Project description:Pseudovibrio a-Proteobacteria have been repeatedly isolated from marine sponges and proposed to be beneficial to the host. Bacterial motility is known to contribute to host colonization. We have previously identified pseudovibriamides A and B, produced in culture by Pseudovibrio brasiliensis Ab134, and shown that pseudovibriamide A promotes flagellar motility. Pseudovibriamides are encoded in a hybrid nonribosomal peptide synthetase-polyketide synthase gene cluster that also includes several accessory genes. Pseudovibriamide A is a linear heptapeptide and pseudovibriamide B is a nonadepsipeptide derived from pseudovibriamide A. Here we define the borders of the pseudovibriamides gene cluster, assign function to biosynthetic genes using reverse genetics and test the hypothesis that pseudovibriamides impact motility by modulating gene transcription. RNA-seq transcriptomic analyses of strains having different compositions of pseudovibriamides suggested that both pseudovibriamides A and B affect genes potentially involved in motility, and that a compensatory mechanism is at play in mutants that produce only pseudovibriamide A, resulting in comparable swarming motility as the wild type. The data gathered suggest that pseudovibriamides A and B have opposite roles in modulating a subset of genes, with pseudovibriamide B having a primary effect in gene activation, and pseudovibriamide A on inhibition. Finally, we observed many differentially expressed genes (up to 29% of the total gene number) indicating that pseudovibriamides have a global effect on transcription that goes beyond motility.
Project description:Animal-algal photosymbioses are a unique group of symbiotic relationships in which animals harbor photosynthetic algae within their cells and tissues. Both marine and freshwater sponges host algal endosymbionts. In previous work, we demonstrated that freshwater sponges can acquire these endosymbionts horizontally through algal infection and that potentially conserved evolutionary pathways may lead to the establishment of the endosymbioses including those involved in endocytosis, ion transport, vesicle-mediated transport, innate immunity, redox regulation, and metabolic processes. Here, we show that algal symbionts can be transferred vertically from algal-bearing overwintering gemmules to adult sponges, and that their proliferation is enhanced by light. Sponges grown under light conditions harbored higher algal loads than those in the dark; however, algae were still able to proliferate and persist in sponges reared in the dark, occupying similar spatial locations to those grown in light. RNA-Seq analysis of algal-bearing sponges across developmental stages in light and dark conditions revealed genetic regulatory pathways involved in the transmission and establishment of the endosymbiosis. Differential expression analysis indicated that the endocytosis and SNARE pathways regulate the internalization and transport of algae at the earliest stage of hatching under light conditions and later in development under dark conditions, potentially contributing to the recruitment of endosymbiotic algae. In sponges hatched in the dark, genes involved in vesicle acidification are regulated, alongside changes in the expression of genes in the pentose phosphate pathway - a key metabolic route involved in redox homeostasis and circadian rhythm regulation via NADPH metabolism, is observed. E. muelleri serves as a versatile model system, supported by robust genomic and transcriptomic resources, for studying host-symbiont interactions. It offers a unique opportunity to investigate the molecular signaling and environmental factors that shape symbiosis in a system where the host can exist with or without algal endosymbionts, symbionts can be acquired either horizontally or vertically, and proliferation of the algae can occur with or without photosynthesis.
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.
