Kirill S. Mironov

Stress Sensors and Signal Transducers in Cyanobacteria

In living cells, the perception of environmental stress and the subsequent transduction of stress signals are primary events in the acclimation to changes in the environment. Some molecular sensors and transducers of environmental stress cannot be identified by traditional and conventional methods. Based on genomic information, a systematic approach has been applied to the solution of this problem in cyanobacteria, involving mutagenesis of potential sensors and signal transducers in combination with DNA microarray analyses for the genome-wide expression of genes. Forty-five genes for the histidine kinases (Hiks), 12 genes for serine-threonine protein kinases (Spks), 42 genes for response regulators (Rres), seven genes for RNA polymerase sigma factors, and nearly 70 genes for transcription factors have been successfully inactivated by targeted mutagenesis in the unicellular cyanobacterium Synechocystis sp. PCC 6803. Screening of mutant libraries by genome-wide DNA microarray analysis under various stress and non-stress conditions has allowed identification of proteins that perceive and transduce signals of environmental stress. Here we summarize recent progress in the identification of sensory and regulatory systems, including Hiks, Rres, Spks, sigma factors, transcription factors, and the role of genomic DNA supercoiling in the regulation of the responses of cyanobacterial cells to various types of stress.

Regulatory role of membrane fluidity in gene expression and physiological functions

Plants, algae, and photosynthetic bacteria experience frequent changes in environment. The ability to survive depends on their capacity to acclimate to such changes. In particular, fluctuations in temperature affect the fluidity of cytoplasmic and thylakoid membranes. The molecular mechanisms responsible for the perception of changes in membrane fluidity have not been fully characterized. However, the understanding of the functions of the individual genes for fatty acid desaturases in cyanobacteria and plants led to the directed mutagenesis of such genes that altered the membrane fluidity of cytoplasmic and thylakoid membranes. Characterization of the photosynthetic properties of the transformed cyanobacteria and higher plants revealed that lipid unsaturation is essential for protection of the photosynthetic machinery against environmental stresses, such as strong light, salt stress, and high and low temperatures. The unsaturation of fatty acids enhances the repair of the damaged photosystem II complex under stress conditions. In this review, we summarize the knowledge on the mechanisms that regulate membrane fluidity, on putative sensors that perceive changes in membrane fluidity, on genes that are involved in acclimation to new sets of environmental conditions, and on the influence of membrane properties on photosynthetic functions.

Light-dependent cold-induced fatty acid unsaturation, changes in membrane fluidity, and alterations in gene expression in Synechocystis ☆

Cold stress causes unsaturation of the membrane lipids. This leads to adjustment of the membrane fluidity, which is necessary for cold acclimation of cells. Here we demonstrate that the cold-induced accumulation of PUFAs in the cyanobacterium Synechocystis is light-dependent. The desA(-)/desD(-) mutant, that lacks the genes for Δ12 and Δ6 desaturases, is still able to adjust the fluidity of its membranes in spite of its inability to synthesize PUFAs and modulate the fatty acid composition of the membrane lipids under cold stress. The expression of cold-induced genes, which are controlled by the cold sensor histidine kinase Hik33, depends on the fluidity of cell membranes and it is regulated by light, though it does not require the activity of the photosynthetic apparatus. The expression of cold-induced genes, which are not controlled by Hik33, does not depend on the membrane fluidity or light. Thus, membrane fluidity determines the temperature dependence of the expression of cold-induced genes that are under control of the Hik33, which might be the sensor of changes in the membrane fluidity. This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial.

Modes of Fatty Acid Desaturation in Cyanobacteria: An Update

Fatty acid composition of individual species of cyanobacteria is conserved and it may be used as a phylogenetic marker. The previously proposed classification system was based solely on biochemical data. Today, new genomic data are available, which support a need to update a previously postulated FA-based classification of cyanobacteria. These changes are necessary in order to adjust and synchronize biochemical, physiological and genomic data, which may help to establish an adequate comprehensive taxonomic system for cyanobacteria in the future. Here, we propose an update to the classification system of cyanobacteria based on their fatty acid composition.

Regulation systems for stress responses in cyanobacteria

The article reviews the main systems that regulate gene expression in cyanobacteria in response to various treatments: low and high temperatures, salt, hyperosmotic and oxidative stresses. The systems for perception of light are also reviewed. Functional characteristics are presented for known two-component regulatory systems, eukaryotic-type serine-threonine protein kinases, σ-subunits of RNA-polymerase, DNA-binding transcription factors. Different mechanisms of perception of stress signals are analyzed, including changes in DNA supercoiling under different stress conditions.

Feedback between fluidity of membranes and transcription of the desB gene for the ω3-desaturase in the cyanobacterium Synechocystis

Cells of prokaryotes (including cyanobacteria) respond to a decrease in the environmental temperature by activation of multiple genes with a low-temperature response. A decrease in temperature causes a reduction of the cell membrane fluidity, which is maintained at an optimum level due to the activity of fatty acid (FA) desaturases. We studied a temperature-dependent expression of the desB gene for the ω3-desaturase in the cyanobacterium Synechocystis, which is able to synthesize polyunsaturated FA, as well as in its double mutant (desA−/desD−), defective in the Δ12- and Δ6-desaturase genes, for which the presence of only monounsaturated FA is typical. During a decrease in temperature, in wild type cells the amount of desB mRNA increased, reaching a maximum value at 24°C. In the desA−/desD− double mutant, an accumulation of the desB transcript was characterized by a maximum at 28–30°C. Thus, using the desB gene encoding the ω3-FA-desaturase, it was demonstrated that a temperature-dependent expression of genes responsible for the maintenance of an optimal fluidity of cell membranes is determined by a physical state of these membranes and is regulated by a feedback mode.

Membrane fluidity controls redox-regulated cold stress responses in cyanobacteria

Membrane fluidity is the important regulator of cellular responses to changing ambient temperature. Bacteria perceive cold by the transmembrane histidine kinases that sense changes in thickness of the cytoplasmic membrane due to its rigidification. In the cyanobacterium Synechocystis, about a half of cold-responsive genes is controlled by the light-dependent transmembrane histidine kinase Hik33, which also partially controls the responses to osmotic, salt, and oxidative stress. This implies the existence of some universal, but yet unknown signal that triggers adaptive gene expression in response to various stressors. Here we selectively probed the components of photosynthetic machinery and functionally characterized the thermodynamics of cyanobacterial photosynthetic membranes with genetically altered fluidity. We show that the rate of oxidation of the quinone pool (PQ), which interacts with both photosynthetic and respiratory electron transport chains, depends on membrane fluidity. Inhibitor-induced stimulation of redox changes in PQ triggers cold-induced gene expression. Thus, the fluidity-dependent changes in the redox state of PQ may universally trigger cellular responses to stressors that affect membrane properties.

Cold-induced gene expression and ω3 fatty acid unsaturation is controlled by red light in Synechocystis

The expression of cold-induced genes, which are controlled by the cold sensor histidine kinase Hik33, and the formation of ω(3) polyunsaturated fatty acids are controlled by light in the cyanobacterium Synechocystis sp. PCC 6803. Cold-induced Hik33-dependent gene expression is initiated by red light (∼700nm), but not by blue or green light. Red light also turns on the ω(3) fatty acid desaturation. Different combinations of other wavelengths in red spectral region (635 and 726nm) had no effect on the red-light-activated cold-induced transcription or fatty acid desaturation. Therefore, the involvement of phytochrome-like photoreceptor(s), similar to phytochromes of higher plants, in this regulation was not confirmed. The absence of light-dependence of gene expression in the mutant cells deficient in Hik33 suggests the involvement of this histidine kinase in direct or mediated with red light regulation of cold responses in Synechocystis.

Aquaporin-deficient mutant of Synechocystis is sensitive to salt and high-light stress

Cyanobacterial aquaporins play an important role in the regulation of various physiological functions: cell volume control, osmotic stress responses, gas exchange. We employed the AqpZ-deficient mutant of Synechocystis to study the role of aquaporins in responses to salt (NaCl) and high light stress. Electron microscopy and paramagnetic resonance revealed that AqpZ-deficient cells are unable to efficiently regulate the cytoplasmic volume under salt stress. Both photosystems (PSII and, especially, PSI) of these cells are more sensitive to NaCl and to high light. Thus, AqpZ of Synechocystis participates in regulation of the photosynthetic activity of PSI and PSII under salt and high-light stress. Our results demonstrate that AqpZ might be necessary for the repair of PSII and PSI after photodamage.

Synechocystis mutants defective in manganese uptake regulatory system, ManSR, are hypersensitive to strong light.

High affinity transport of manganese ions (Mn2+) in cyanobacteria is carried by an ABC-type transporter, encoded by the mntCAB operon, which is derepressed by the deficiency of Mn2+. Transcription of this operon is negatively regulated by the two-component system consisting of a sensory histidine kinase ManS and DNA-binding response regulator ManR. In this study, we examined two Synechocystis mutants, defective in ManS and ManR. These mutants were unable to grow on high concentrations of manganese. Furthermore, they were sensitive to high light intensity and unable to recover after short-term photoinhibition. Under standard illumination and Mn2+ concentration, mutant cells revealed the elevated levels of transcripts of genes involved in the formation of Photosystem II (psbA, psbD, psbC, pap-operon). This finding suggests that, in mutant cells, the PSII is sensitive to high concentrations of Mn2+ even at relatively low light intensity.