Expressed 2-5A synthetase genes and pseudogenes in the marine sponge Geodia barretti.

The 2',5'-oligoadenylate synthetases (2-5A synthetases, OAS) form a family of proteins presented in many branches of Metazoa. The phylum Porifera (sponges) contains OAS proteins which are different from those in vertebrates and form a distinct OAS subfamily. In turn, OAS proteins from different genera of Demospongia show rather low similarities in their primary structures. To ascertain divergence of the OAS genes within a particular sponge genus, we identified the OAS genes from the marine sponge Geodia barretti and compared them with those from another member of the genus Geodia, Geodia cydonium. The identity and similarity of the OAS sequences found in G. barretti with those from G. cydonium were considerably higher than identities and similarities compared with those from other sponges, 75% and 85% versus 27-30% and 42-47%, respectively. We also established the presence of a transcriptionally active polymorphic OAS pseudogene in the genome of G. barretti. The transcripts of the OAS pseudogene(s) lack several internal exons encoding necessary motifs for OAS enzymatic activity. The maintenance and further diversification of OAS gene(s) and pseudogene(s) suggest the prevalence of gene duplication events over the loss of gene duplicates in Geodia genomes during the evolution.

Cytoskeleton alterations induced by Geodia corticostylifera depsipeptides in breast cancer cells

Crude extracts of the marine sponge Geodia corticostylifera from Brazilian Coast have previously shown antibacterial, antifungal, cytotoxic, haemolytic and neurotoxic activities. The present work describes the isolation of the cyclic peptides geodiamolides A, B, H and I (1–4) from G. corticostylifera and their anti-proliferative effects against sea urchin eggs and human breast cancer cell lineages. Its structure–activity relationship is discussed as well. In an initial series of experiments these peptides inhibited the first cleavage of sea urchin eggs (Lytechinus variegatus). Duplication of nuclei without complete egg cell division indicated the mechanism of action might be related to microfilament disruption. Further studies showed that the geodiamolides have anti-proliferative activity against human breast cancer cell lines (T47D and MCF7). Using fluorescence techniques and confocal microscopy, we found evidence that the geodiamolides A, B, H and I act by disorganizing actin filaments of T47D and MCF7 cancer cells, in a way similar to other depsipeptides (such as jaspamide 5 and dolastatins), keeping the normal microtubule organization. Normal cells lines (primary culture human fibroblasts and BRL3A rat liver epithelial cells) were not affected by the treatment as tumor cells were, thus indicating the biomedical potential of these compounds.

Neurotoxic activity induced by a haemolytic substance in the extract of the marine sponge Geodia corticostylifera

In our search for marine bioactive compounds we chose a Brazilian Coast sponge, Geodia corticostylifera (Demospongiae), whose extracts showed previously antibacterial and antifungal activities. In the present work we studied the following toxic properties of G. corticostylifera extract: neurotoxic (in mouse neuromuscular junction); mouse acute toxicity (IP) and haemolytic (against mouse and frog erythrocytes). Insertion of ionic channels in planar lipid bilayers in presence of a haemolytic purified fraction of the extract was observed. The toxic activities of G. corticostylifera crude extract are related to the formation of ionic pores in the cell membrane, which induce the release of haemoglobin from erythrocytes, and depolarization of nerve and muscle membranes. These last physiological effects cause the blockade of the diaphragm contractions, leading to death through respiratory arrest.