Transcriptional Control of Photosynthesis Genes: The Evolutionarily Conserved Regulatory Mechanism in Plastid Genome Function

Chloroplast sensor kinase (CSK) is a bacterial-type sensor histidine kinase found in chloroplasts—photosynthetic plastids—in eukaryotic plants and algae. Using a yeast two-hybrid screen, we demonstrate recognition and interactions between: CSK, plastid transcription kinase (PTK), and a bacterial-type RNA polymerase sigma factor-1 (SIG-1). CSK interacts with itself, with SIG-1, and with PTK. PTK also interacts directly with SIG-1. PTK has previously been shown to catalyze phosphorylation of plastid-encoded RNA polymerase (PEP), suppressing plastid transcription nonspecifically. Phospho-PTK is inactive as a PEP kinase. Here, we propose that phospho-CSK acts as a PTK kinase, releasing PTK repression of chloroplast transcription, while CSK also acts as a SIG-1 kinase, blocking transcription specifically at the gene promoter of chloroplast photosystem I. Oxidation of the photosynthetic electron carrier plastoquinone triggers phosphorylation of CSK, inducing chloroplast photosystem II while suppressing photosystem I. CSK places photosystem gene transcription under the control of photosynthetic electron transport. This redox signaling pathway has its origin in cyanobacteria, photosynthetic prokaryotes from which chloroplasts evolved. The persistence of this mechanism in cytoplasmic organelles of photosynthetic eukaryotes is in precise agreement with the CoRR hypothesis for the function of organellar genomes: the plastid genome and its primary gene products are Co-located for Redox Regulation. Genes are retained in plastids primarily in order for their expression to be subject to this rapid and robust redox regulatory transcriptional control mechanism, whereas plastid genes also encode genetic system components, such as some ribosomal proteins and RNAs, that exist in order to support this primary, redox regulatory control of photosynthesis genes. Plastid genome function permits adaptation of the photosynthetic apparatus to changing environmental conditions of light quantity and quality

Subcellular distribution of glutathione and cysteine in cyanobacteria

Glutathione plays numerous important functions in eukaryotic and prokaryotic cells. Whereas it can be found in virtually all eukaryotic cells, its production in prokaryotes is restricted to cyanobacteria and proteobacteria and a few strains of gram-positive bacteria. In bacteria, it is involved in the protection against reactive oxygen species (ROS), osmotic shock, acidic conditions, toxic chemicals, and heavy metals. Glutathione synthesis in bacteria takes place in two steps out of cysteine, glutamate, and glycine. Cysteine is the limiting factor for glutathione biosynthesis which can be especially crucial for cyanobacteria, which rely on both the sufficient sulfur supply from the growth media and on the protection of glutathione against ROS that are produced during photosynthesis. In this study, we report a method that allows detection and visualization of the subcellular distribution of glutathione in Synechocystis sp. This method is based on immunogold cytochemistry with glutathione and cysteine antisera and computer-supported transmission electron microscopy. Labeling of glutathione and cysteine was restricted to the cytosol and interthylakoidal spaces. Glutathione and cysteine could not be detected in carboxysomes, cyanophycin granules, cell walls, intrathylakoidal spaces, periplasm, and vacuoles. The accuracy of the glutathione and cysteine labeling is supported by two observations. First, preadsorption of the antiglutathione and anticysteine antisera with glutathione and cysteine, respectively, reduced the density of the gold particles to background levels. Second, labeling of glutathione and cysteine was strongly decreased by 98.5% and 100%, respectively, in Synechocystis sp. cells grown on media without sulfur. This study indicates a strong similarity of the subcellular distribution of glutathione and cysteine in cyanobacteria and plastids of plants and provides a deeper insight into glutathione metabolism in bacteria

Numerical modelling of nonlinear extreme waves in the presence of wind

A numerical wave flume with fully nonlinear free surface boundary conditions is adopted to investigate the temporal characteristics of extreme waves in the presence of wind at various speeds. Incident wave trains are numerically generated by a piston-type wave maker, and the wind-excited pressure is introduced into dynamic boundary conditions using a pressure distribution over steep crests, as defined by Jeffreys’ sheltering mechanism. A boundary value problem is solved by a higher-order boundary element method (HOBEM) and a mixed Eulerian-Lagrangian time marching scheme. The proposed model is validated through comparison with published experimental data from a focused wave group. The influence of wind on extreme wave properties, including maximum extreme wave crest, focal position shift, and spectrum evolution, is also studied. To consider the effects of the wind-driven currents on a wave evolution, the simulations assume a uniform current over varying water depth. The results show that wind causes weak increases in the extreme wave crest, and makes the nonlinear energy transfer non-reversible in the focusing and defocusing processes. The numerical results also provide a comparison to demonstrate the shifts at focal points, considering the combined effects of the winds and the wind-driven currents

Comparison of novel ladle slag treatment and handling systems based on resource-efficiency metrics

Ladle slag is a by-product common to electric and basic oxygen steelmaking furnaces which is gaining increasing attention as a secondary material. Its main recycling path is internal to steelmaking process, since it can replace the lime used to remove impurities. However, storing and handling slag for internal recycling is problematic because cooled ladle slag soon becomes extremely dusty, determining harsh environmental conditions at the plant. Recently, a novel solution based on granulation of ladle slag was presented on the market, which could be integrated in the steelmaking process using diverse handling and storage systems. The implementation of such systems requires resources, specifically energy, but may produce benefits such as lower pollution from particulate emissions and easier storage, leading to lower material losses, reduced landfill disposal and savings of primary mineral resources. In this paper, three alternative treatment and handling systems are analyzed and ranked using ad hoc defined first level resource efficiency metrics. Results show that the best alternative in terms of carbon emission intensity is the more advanced configuration, which includes granulation within a casing and automatic transport with apron conveyors; however, open granulation with current handling systems apparently minimizes primary energy intensity. A possible cause for this discrepancy is that emission factors and primary energy consumption factors obtained from official sources refer to different years, and hence to a different electric energy generation mix. A clear ranking between the basic and the most advanced configuration cannot be obtained, but the resource efficiency evaluation leads to exclude the intermediate configuration (granulation within a casing and traditional materials handling) which is apparently dominated by the remaining alternatives

Multiple evidence for nucleotide metabolism in the chloroplast thylakoid lumen

The apparatus of photosynthetic energy conversion in chloroplasts is quite well characterized with respect to structure and function. Light-driven electron transport in the thylakoid membrane is coupled to synthesis of ATP, used to drive energy-dependent metabolic processes in the stroma and the outer surface of the thylakoid membrane. The role of the inner (luminal) compartment of the thylakoids has, however, remained largely unknown although recent proteomic analyses have revealed the presence of up to 80 different proteins. Further, there are no reports concerning the presence of nucleotides in the thylakoid lumen. Here, we bring three lines of experimental evidence for nucleotide-dependent processes in this chloroplast compartment. (i) The thylakoid lumen contains a protein of 17.2 kDa, catalyzing the transfer of the γ-phosphate group from ATP to GDP, proposed to correspond to the nucleoside diphosphate kinase III. (ii) The 33-kDa subunit of photosystem II, bound to the luminal side of the thylakoid membrane and associated with the water-splitting process, can bind GTP. (iii) The thylakoid membrane contains a nucleotide transport system that is suggested to be associated with a 36.5-kDa nucleotide-binding protein. Our results imply, against current dogmas, that the thylakoid lumen contains nucleotides, thereby providing unexpected aspects on this chloroplast compartment from a metabolic and regulatory perspective and expanding its functional significance beyond a pure bioenergetic function