Project description:Harmful algal blooms present severe environmental threats, impacting water quality, aquatic ecosystems, and human health. The frequency and intensity of these blooms are rising, largely driven by global warming and changing climatic conditions. There is an urgent need for innovative methods to monitor blue-green algae, also known as cyanobacteria, to enable the implementation of preventative measures. Here, we show that native mass spectrometry is an effective tool for detecting cyanobacteria directly from lake samples, both prior and during bloom formation. Our approach allows for the rapid characterization of cyanobacterial populations within lakes, offering valuable insights into the dynamics of cyanobacterial species associated with harmful algae blooms. Overall, we highlight the exceptional capability of native mass spectrometry in directly detecting and monitoring cyanobacterial blooms, which will support the development of more effective strategies to mitigate this growing environmental challenge.
Project description:Cyanobacteria are photosynthetic prokaryotes vital to global biogeochemical cycles. However, some form harmful algal blooms (cyanoHABs) that disrupt ecosystems and produce toxins. The mechanisms by which these blooms form have yet to be fully understood, particularly the role of extracellular components. Here, we present a 2.4 Å cryo-EM structure of a pilus, termed the cyanobacterial tubular (CT) pilus, found in the cyanoHAB-forming species Microcystis aeruginosa. The pilin exhibits a novel protein fold, forming a tubular pilus structure with tight, double-layer anti-parallel β-sheet interactions. We show that CT pili are essential for buoyancy by facilitating the formation of micro-colonies, which increases drag force and prevents sinking. Furthermore, the CT pilus surface is heavily glycosylated with ten monosaccharide modifications per pilin. We also found that CT pili can enrich microcystin, potentially enhancing cellular resilience, and co-localize with iron-enriched extracellular matrix components. Thus, we propose that this pilus plays an important role in the proliferation of cyanoHABs. This newly discovered pilus family appears to be widely distributed across several cyanobacterial orders, including Chroococcales, Nostocales, and Synechococcales. Our structural and functional characterization of CT pili provide insights into cyanobacterial cell morphology, physiology, and toxin interactions, and identify potential targets for disrupting bloom formation.
Project description:Cyanobacteria produce various cyanotoxins, which can cause severe effects to other organisms. Microcystins, one group of such toxins, primarily produced by species of Microcystis, are strong hepatotoxins and inhibit potently protein phosphatases 1 and 2A. Microcystin is the most studied cyanotoxin, however, others are not investigated. Eutrophication of water bodies promotes the occurrence of toxic algal blooms and since a anthropogenic caused increase in eutrophication events can be observed, it is becoming increasingly important to study the consequences and to increase the knowledge on toxins associated with algal blooms. Recently a new cyanobacteria toxin from a Microcystis strain, CP1020, was described. CP1020 belongs to the class of cyanopeptolins and its toxicity was shown to be comparable to that of microcystin (Gademann et al., 2009). It is a strong protease inhibitor inhibiting trypsin in the picomolar range (IC50 = 670 pM) and effects survival of the freshwater crustacean Thamnocephalus platyurus (LC50) 8.8 μM (Gademann et al., 2009). Nothing is known, however, about the toxicity of CP1020 to fish. Furthermore, no information is available on the toxic modes of action, in addition to the proteinase activity. Consequently our study has the aim to elucidate the modes of action of CP1020 on zebrafish eleuthero-embryos. By using a microarray technique, we will analyse alterations of global gene expression by CP1020 at two different concentrations. Thereby, we hope to elucidate the whole array of affected biological pathways to elucidate the mechanisms by which CP1020 affect fish.
Project description:Cyanobacteria produce various cyanotoxins, which can cause severe effects to other organisms. Microcystins, one group of such toxins, primarily produced by species of Microcystis, are strong hepatotoxins and inhibit potently protein phosphatases 1 and 2A. Microcystin is the most studied cyanotoxin, however, others are not investigated. Eutrophication of water bodies promotes the occurrence of toxic algal blooms and since a anthropogenic caused increase in eutrophication events can be observed, it is becoming increasingly important to study the consequences and to increase the knowledge on toxins associated with algal blooms. Recently a new cyanobacteria toxin from a Microcystis strain, CP1020, was described. CP1020 belongs to the class of cyanopeptolins and its toxicity was shown to be comparable to that of microcystin (Gademann et al., 2009). It is a strong protease inhibitor inhibiting trypsin in the picomolar range (IC50 = 670 pM) and effects survival of the freshwater crustacean Thamnocephalus platyurus (LC50) 8.8 M-NM-<M (Gademann et al., 2009). Nothing is known, however, about the toxicity of CP1020 to fish. Furthermore, no information is available on the toxic modes of action, in addition to the proteinase activity. Consequently our study has the aim to elucidate the modes of action of CP1020 on zebrafish eleuthero-embryos. By using a microarray technique, we will analyse alterations of global gene expression by CP1020 at two different concentrations. Thereby, we hope to elucidate the whole array of affected biological pathways to elucidate the mechanisms by which CP1020 affect fish. Gene expression in zebrafish eleuthero-embryos was measured after exposure for 96h to 100 ug/L and 1000 ug/L CP1020 or to the respective controls. A total of 12 arrays (Agilent 4 M-CM-^W 44 K Zebrafish microarray) were used, including four for the solvent control group, four for the 100 M-NM-<g/L and four for the 1000 M-NM-<g/L CP1020 dose group.