Microcystins are cyanobacterial toxins that represent a serious threat to drinking water and recreational lakes worldwide. Here, we show that microcystin fulfils an important function within cells of its natural producer Microcystis. The microcystin deficient mutant ΔmcyB showed significant changes in the accumulation of proteins, including several enzymes of the Calvin cycle, phycobiliproteins and two NADPH-dependent reductases. We have discovered that microcystin binds to a number of these proteins in vivo and that the binding is strongly enhanced under high light and oxidative stress conditions. The nature of this binding was studied using extracts of a microcystin-deficient mutant in vitro. The data obtained provided clear evidence for a covalent interaction of the toxin with cysteine residues of proteins. A detailed investigation of one of the binding partners, the large subunit of RubisCO showed a lower susceptibility to proteases in the presence of microcystin in the wild type. Finally, the mutant defective in microcystin production exhibited a clearly increased sensitivity under high light conditions and after hydrogen peroxide treatment. Taken together, our data suggest a protein-modulating role for microcystin within the producing cell, which represents a new addition to the catalogue of functions that have been discussed for microbial secondary metabolites.
Cyanobacteria are versatile unicellular phototrophic microorganisms that are highly abundant in many environments. Owing to their capability to utilize solar energy and atmospheric carbon dioxide for growth, cyanobacteria are increasingly recognized as a prolific resource for the synthesis of valuable chemicals and various biofuels. To fully harness the metabolic capabilities of cyanobacteria necessitates an in-depth understanding of the metabolic interconversions taking place during phototrophic growth, as provided by genome-scale reconstructions of microbial organisms. Here we present an extended reconstruction and analysis of the metabolic network of the unicellular cyanobacterium Synechocystis sp. PCC 6803. Building upon several recent reconstructions of cyanobacterial metabolism, unclear reaction steps are experimentally validated and the functional consequences of unknown or dissenting pathway topologies are discussed. The updated model integrates novel results with respect to the cyanobacterial TCA cycle, an alleged glyoxylate shunt, and the role of photorespiration in cellular growth. Going beyond conventional flux-balance analysis, we extend the computational analysis to diurnal light/dark cycles of cyanobacterial metabolism.
The biosynthesis of glycogen or starch is one of the main strategies developed by living organisms for the intracellular storage of carbon and energy. In phototrophic organisms, such polyglucans accumulate due to carbon fixation during photosynthesis and are used to provide maintenance energy for cell integrity, function and viability in dark periods. Moreover, it is assumed that glycogen enables cyanobacteria to cope with transient starvation conditions, as observed in most micro-organisms. Here, glycogen accumulates when an appropriate carbon source is available in sufficient amounts but growth is inhibited by lack of other nutrients. In this study, the role of glycogen in energy and carbon metabolism of phototrophic cyanobacteria was first analysed via a comparative physiological and metabolic characterization of knockout mutants defective in glycogen synthesis. We first proved the role of glycogen as a respiratory substrate in periods of darkness, the role of glycogen as a reserve to survive starvation periods such as nitrogen depletion and the role of glycogen synthesis as an ameliorator of carbon excess conditions in the model organism Synechocystis sp. PCC 6803. We provide striking new insights into the complex carbon and nitrogen metabolism of non-diazotrophic cyanobacteria: a phenotype of sensitivity to photomixotrophic conditions and of reduced glucose uptake, a non-bleaching phenotype based on an impaired acclimation response to nitrogen depletion and furthermore a phenotype of energy spilling. This study shows that the analysis of deficiencies in glycogen metabolism is a valuable tool for the identification of metabolic regulatory principles and signals.
and Manchester Interdisciplinary Biocentre, University of Manchester, M1 7DN Manchester, United Kingdom (R.S.) Unicellular cyanobacteria have attracted growing attention as potential host organisms for the production of valuable organic products and provide an ideal model to understand oxygenic photosynthesis and phototrophic metabolism. To obtain insight into the functional properties of phototrophic growth, we present a detailed reconstruction of the primary metabolic network of the autotrophic prokaryote Synechocystis sp. PCC 6803. The reconstruction is based on multiple data sources and extensive manual curation and significantly extends currently available repositories of cyanobacterial metabolism. A systematic functional analysis, utilizing the framework of flux-balance analysis, allows the prediction of essential metabolic pathways and reactions and allows the identification of inconsistencies in the current annotation. As a counterintuitive result, our computational model indicates that photorespiration is beneficial to achieve optimal growth rates. The reconstruction process highlights several obstacles currently encountered in the context of large-scale reconstructions of metabolic networks.Cyanobacteria are among the evolutionarily oldest organisms and are the only known prokaryotes capable of plant-like oxygenic photosynthesis. As primary producers in aquatic environments, they play an important role in global CO 2 assimilation and oxygen recycling. Recently, cyanobacteria have also attracted growing attention for economic purposes, including drug discovery and as prolific producers of natural products (Sielaff et al., 2006;Tan, 2007). In particular their ability to directly convert atmospheric CO 2 into biomass and organic compounds, driven by sunlight, offers considerable potential as a novel and renewable resource for bioenergy (Deng and Coleman, 1999;Atsumi et al., 2009;Mascarelli, 2009;Lindberg et al., 2010).Among the diverse cyanobacterial strains, Synechocystis sp. PCC 6803 is one of the most extensively studied model organisms for the analysis of photosynthetic processes. With a rich compendium of genomic, biochemical, and physiological data available, Synechocystis sp. PCC 6803, therefore, offers an ideal starting point to obtain insights into the systemic properties of phototrophic metabolism. The prerequisite for such a systemic description is a detailed reconstruction of the metabolic network of the organism: that is, a reconstruction of the comprehensive set of enzyme-catalyzed reactions required to support cellular growth and maintenance. Once a metabolic reconstruction is available, the vast array of methods developed by computational systems biology over the past decades allows us to dissect the functioning and interplay of possible metabolic routes and biochemical interconversions. In this respect, constraint-based modeling, most notably flux-balance analysis (FBA), has become a quasi-standard in the field. FBA is increasingly utilized to elucidate and characterize largescale network...
Background: The colonial cyanobacterium Microcystis proliferates in a wide range of freshwater ecosystems and is exposed to changing environmental factors during its life cycle. Microcystis blooms are often toxic, potentially fatal to animals and humans, and may cause environmental problems. There has been little investigation of the genomics of these cyanobacteria.
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